BioQuakes

AP Biology class blog for discussing current research in Biology

Tag: Gene-editing (Page 1 of 2)

Can Technology Ketchup To These Super Tomatoes?

Sicilian Rouge tomatoes are one of the first foods made with CRISPR-Cas9 technology to be sold to the public. An article by Emily Waltz, of Scientific American, goes in depth on how these tomatoes are taking Japan by storm. Sanatech Seed, a company based in Tokyo, has edited the tomatoes to have a large amount of GABA(γ-aminobutyric acid).  According to the company, GABA supposedly lowers blood pressure and promotes relaxation when ingested orally.

In Japan, GABA is a popular addition to many foods, drinks and other products such as chocolates. Hiroshi Ezra works as both the chief technology officer at Sanatech and a plant molecular biologist at the University of Tsukuba. He says that “GABA is a famous health-promoting compound in Japan. It’s like vitamin C…That’s why we chose this as our first target for our genome editing technology. “

 

CRISPR has been used in a myriad of ways by plant bioengineers. Non-browning mushrooms and drought-tolerant soybeans are just a few examples of this. However, Sanatech’s Sicilian Rouge tomato was the first CRISPR-edited food known to be commercialized.

 

But what is CRISPR and why has it become so popular? Yourgenome.org effectively explains what the different parts of the CRISPR-Cas9 technology do. The system is made of two parts: the enzyme and RNA. The enzyme is called Cas9 and its role in gene editing is to ‘cut’ the specific genome in strand of DNA so that the mutation can be made. The RNA acts as a guide for the enzyme, which is why it is called gRNA. The piece of RNA is made of an approximately 20 base sequence that is a part of the longer RNA ‘scaffold’. When the strand binds to the DNA the 20 base sequence guides the Cas9 to the part of the genome that is meant to be cut. The scaffold is able to find the correct genome because its bases are made to be specifically complementary to the target genome. Once the genome is cut the cell recognizes the cut in the DNA and repairs it. It is when this repair takes place that the changes/mutations to the genome occur. 

4.3. The CRISPR Cas 9 system III

The processes of CRISPR are similar to what we learned about in biology too. During DNA replication, small complementary strands of RNA act as primers so that DNA polymerase can add to anc continue the chain. DNA polymerase also ‘proofreads’ strands of DNA for any mistakes which it would cut out and replace with the correct nucleotides. The Ligase then reforms the phosphodiester bonds which hold the nucleotides together. This process of error correction is what takes place once the Cas9 cuts the genomes.

 

Another type of DNA editing is called TALENs or transcription activator-like effector nucleases. A company called Calyxt commercialized TALENs through their genetically edited soybean oil that is free of trans fats. Gene editing hasn’t only been bound to plants, but also animals too. In October of last year Japan approved CRISPR two gene-edited fish. One was an edited tiger puffer which “exhibits depressed appetite suppression”. The other was a Red Sea bream which was edited to have “increased muscle growth”.

 

From super-crops to super-fish, it appears as though there are no limits for CRISPR in our daily lives. It’s amazing how precise technology has allowed us to alter the nutrition of the food we eat. I wonder what other possibilities lie in the future of CRISPR and how they will affect our society.

Unnatural Selection: The Future of The Future?

Imagine it’s Saturday night, you are snowed in until the morning and you need a way to pass the time. Like many people, you resort to Netflix. Upon browsing through the vast selection of horror, comedy, and romantic films, you decide you are in the mood for a documentary. Scrolling through the options, you stop at a title that grabs your attention: Unnatural Selection.

Since you are an AP Biology student, you immediately connect the words “Natural Selection” to the work of Charles Darwin, the study of genetics, and most importantly: evolution. In brief, natural selection is the survival and reproduction of the fittest, the idea that organisms with traits better suited to living in a specific environment will survive to reproduce offspring with similar traits. Those with unfavorable traits may not be able to reproduce, and therefore those traits are no longer passed down through that species. Natural selection is a mechanism for genetic diversity in evolution, and it is how species adapt to certain environments over many generations.

If genetic diversity enables natural selection, then what enables unnatural selection? Well, If natural selection eradicates unfavorable traits naturally, then unnatural selection essentially eradicates unfavorable traits or promotes favorable traits artificially.

The Netflix docuseries “Unnatural Selection” focuses on the emergence of a new gene-editing technology named CRISPR (an acronym for “Clustered regularly interspaced short palindromic repeats”). CRISPR is a revolutionary new method of DNA editing, which could help cure both patients with genetic diseases and patients who are at risk of inheriting unwanted genetic diseases. The two pioneers of this technology, Emmanuelle Charpentier and Jennifer Doudna, recently won Nobel Prizes in Chemistry for their work on CRISPR.

CRISPR illustration gif animation 1

Animation of CRISPR using guide RNA to identify where to cut the DNA, and cutting the DNA using the Cas9 enzyme

CRISPR works with the Cas9 enzyme to locate and cut a specific segment of DNA. Scientists first identify the sequence of the human genome, and locates a specific region that needs to be altered. Using that sequence, they design a guide RNA strand that will help the Cas9 enzyme, otherwise known as the “molecular scissors” to locate the specific gene, and then make precision cuts. With the affected region removed, scientists can now insert a correct sequence in its place.

Using the bacterial quirk that is CRISPR, scientists have essentially given anyone with a micropipette and an internet connection the power to manipulate the genetic code of any living thing.

Megan Molteni / WIRED

CRISPR is just the beginning of gene editing, introducing a new field of potential gene editing research and applications. But with great power comes great responsibility — and great controversy. Aside from the obvious concerns, people speculating the safety, research, and trials of this new treatment, CRISPR headlines are dominated by a hefty ethical dilemma. On one hand, treating a patient for sickle cell anemia will rid them of pain and suffering, and allows their offspring to enjoy a normal life as well. However, by eliminating the passing down of this trait, sickle cell anemia is slowly eliminated from the human gene pool. Rather than natural selection choosing the path of human evolution — we are. We are selecting which traits we deem “abnormal” and are removing them scientifically. Although CRISPR treatment is intended to help people enjoy normal lives and have equally as happy children, putting evolution into the hands of those evolving can result in more drastic effects in the future.

For our generation, CRISPR seems like a magical cure for genetic diseases. But for future generations, CRISPR could very well be seen as the source of many problems such as overpopulation, low genetic diversity, and future alterations such as “designer babies.” Humans have reached the point where we are capable of our future. Is CRISPR going to solve all of our problems, or put an end to the diverse human race as we know it? Comment how you feel down in the comments.

 

CRISP[ie]R Corn Kernels?

Corn is unique in the way that its genome is highly complex, thus causing it to be very difficult to edit those genes with technology such as CRISPR. CRISPR is an advanced technology that is used to find a specific portion of DNA in a cell and then it alters that piece of DNA. To learn more about CRISPR, click here.

CRISPR CAS9 technology

In a recent study at Cold Spring Harbor Laboratory, researchers attempted to modify the growth of stem cells and promotor regions in corn using CRISPR. Thousands of years ago, corn was just a plant covered in weeds that formed very few kernels on its surface. Through gene editing technologies, scientists were able to transform the hopeless plant into a delicious vegetable with juicer kernels bursting from all surfaces. To increase the number of corn kernels 0n the surface of the plant, Professor David Jackson along with Lei Liu worked in collaboration with Professor Madelaine Bartlett from the University of Massachusetts Amherst. They were one of the first groups to tackle the editing of corn’s complex set of DNA.

Zea mays 'Ottofile giallo Tortonese' MHNT.BOT.2015.34.1

We are currently learning in AP Biology how DNA is replicated and can be altered. In replication, DNA is first untwisted by a helicase enzyme. Similarly, CRISPR uses an enzyme called Cas9 that unzips the DNA. This allows for the newly created strand of RNA to be matched to the target DNA. The Cas9 then cuts the DNA strand which causes the cell to attempt and put the strand back together and this results in new genes being formed because the DNA sequence is altered. This is just like how in replication, the DNA polymerase adds nucleotides to an existing strand of DNA. This video also provides a great visual description of how CRISPR can edit existing genes.

Since corn is a plant, it consists of plant cells that have a much stronger cell wall than animal cells do. This makes it harder for the CRISPR to access the cell’s DNA and make edits. CRISPR can be used to disrupt genes and eliminate them, as well as help the promoter regions which activate the genes instead. Corn kernel development depends on the genes supporting stem cell growth. They experimented by targeting random areas of the promoter to see which part will change the number of kernels on the cob.

Ontario-Corn-field 03

As a veggie-lover myself, I am so glad that these new gene-editing procedures allow for fuller, juicier corn kernels. Not only is this beneficial to those who eat corn on the cob or choose to enjoy a moist slice of cornbread, but also to those who love to sit down with a big bowl of popcorn to watch a movie. If a vegetable with such complex genes as corn is able to be improved, imagine what the future holds for other plants yielding yummy additions to our diets!

Redesigned Cas9 protein provides safer gene editing than ever before!

Gene editing is a group of technologies that give scientists the ability to change an organism’s DNA. These technologies allow genetic material to be added, removed, or altered at particular locations in the genome. Several approaches to genome editing have been developed. A recent one is known as CRISPR-Cas9, which is short for clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9. The CRISPR-Cas9 system has generated a lot of excitement in the scientific community because it is faster, cheaper, more accurate, and more efficient than other existing genome editing methods.

One of the challenges that come using CRISPR-based gene editing within humans is that the molecular machinery may sometimes make edits to the wrong section of a host’s genome. This is problematic because it creates the possibility that an attempt to repair a genetic mutation in one location in the genome could accidentally create a dangerous new mutation in another spot. Scientists at The University of Texas at Austin have redesigned a key component of a widely used CRISPR-based gene-editing tool, called Cas9, to be thousands of times less likely to target the wrong stretch of DNA while remaining just as efficient as the original version, making it potentially much safer.

The CRISPR-Cas9 system works similarly in the lab. Researchers create a small piece of RNA with a short ‘guide’ sequence that binds to a specific target sequence of DNA in a genome. The RNA also binds to the Cas9 enzyme. As in bacteria, the modified RNA is used to recognize the DNA sequence, and the Cas9 enzyme cuts the DNA at the targeted location. Although Cas9 is the enzyme that is used most often, other enzymes can also be used. Once the DNA is cut, researchers use the cell’s own DNA repair machinery to add or delete pieces of genetic material, or to make changes to the DNA by replacing an existing segment with a customized DNA sequence.

Other labs have redesigned Cas9 to reduce off-target interactions, but so far, all these versions improve accuracy by sacrificing speed. SuperFi-Cas9, as this new version has been named, is 4,000 times less likely to cut off-target sites but just as fast as naturally occurring Cas9. Scientists say you can think of the different lab-generated versions of Cas9 as different models of self-driving cars. Most models are really safe, but they have a top speed of 10 miles per hour.

In my opinion, setting aside any and all ethical concerns, genome editing is of great interest in the prevention and treatment of human diseases. Currently, most research on genome editing is done to understand diseases using cells and animal models. Scientists are still working to determine whether this approach is safe and effective for use in people. It is being explored in research on a wide variety of diseases, including single-gene disorders such as cystic fibrosis, hemophilia, and sickle cell disease. It also holds promise for the treatment and prevention of more complex diseases, such as cancer, heart disease, mental illness, and human immunodeficiency virus (HIV) infection.

How Does Activation of p53 Effect the Use of CRISPR?

In a study conducted at Karolinska Institutet in Sweden, researchers looked into CRISPR gene editing and how that can play a critical role in mutated cancer cells as well as the medical field. CRISPR is “programmed to target specific stretches of genetic coding and to edit DNA at the precise location;” specifically, the CRISPR system binds to the DNA and cuts it, therefore, shutting the targeted gene off. Researchers can also permanently alter genes in living cells and organisms, and in the future, using this method they may even be used to treat genetic causes of diseases. Although CRISPR sounds amazing, will it really be as great as it seems?

CRISPR CAS9 technology

CRISPR

There are a few obstacles that need to be overcome before CRISPR can even become regularly administered in hospitals. The first is to understand how cells will behave once they are subjected to DNA damage which is caused by CRISPR in a controlled manner. When cells are damaged they activate a protein called p53 which has negative and positive effects on the procedure. The technique is less effective when p53 is activated, however, when p53 is not activated cells can grow uncontrollably and become cancerous. Cells, where p53 is not activated, have a higher survival rate when subjected to CRISPR and because of this can accumulate in mixed cell populations. Researchers have also found a network of linked genes that have a similar effect to p53 mutations, so inhibiting p53 also prevents these cells from mutating. 

Long Jiang, a doctoral student at the Department of Medicine at Karolinska Institutet, says that “it can be contrary to inhibit p53 in a CRISPR context. However, some literature supports the idea that p53 inhibition can make CRISPR more effective.” By doing this it can also counteract the replication of cells with mutations in p53 as well as genes that are associated with the mutations. This research established a network of possible genes that should be carefully controlled for mutations during CRISPR. This will hopefully allow for mutations to be regulated and contained more efficiently.

DNA, or deoxyribonucleic acid, is a long molecule that contains a genetic code; “like a recipe book it holds all the instructions for making the proteins in our bodies.” Most DNA is found in the nucleus of the cell, but a small amount can also be found in the mitochondria. DNA is a key part of reproduction because genetic heredity comes from the passing down of DNA from parents to offspring. Altering this DNA can have an impact on a number of someone’s physical characteristics. CRISPR does just that. It can be used to edit genes by finding a specific piece of DNA inside a cell and then modifying it. Since CRISPR is so new, it has its positives and negatives, but overall it is a groundbreaking discovery. 

DNA double helix horizontal

DNA

In conclusion, even though cells seem to gain p53 mutations from CRISPR, it has been discovered that most of the cell mutations were there from the start. Even though this is still an issue, we don’t know to what extent it can cause greater harm, so it will be exciting to see the new discoveries in the future!

CRISPR Gene Editing: The Future of Food?

Biology class has taught me a lot about genes and DNA – I know genes code for certain traits, DNA is the code that makes up genes, and that genes are found on chromosomes. I could even tell two parents, with enough information, the probabilities of different eye colors in their children! However, even with all this information, when I first heard “gene editing technology,” I thought, “parents editing what their children will look like,” and while this may be encapsulated in the CRISPR gene editing technology, it is far from its purpose! So, if you’re like me when I first started my CRISPR research, you have a lot to learn! Let’s dive right in!

CRISPR

Firstly, what is CRISPR Gene Editing? It is a genetic engineering technique that “edits genes by precisely cutting DNA and then letting natural DNA repair processes to take over” (http://www.crisprtx.com/gene-editing/crispr-cas9).  Depending on the cut of DNA, three different genetic edits can occur: if a single cut in the DNA is made, a gene can be inactivated; if two separate DNA sites are cut, the middle part of DNA will be deleted, and the separate cuts will join together; and if the same two separate pieces of DNA are cut, but a DNA template is added, the middle part of DNA that would have been deleted can either be corrected or completely replaced. This technology allows for endless possibilities of advancements, from reducing toxic protein to fighting cancer, due to the countless ways it can be applied. Check out this link for some other incredible ways to apply CRISPR technology!

In this blog post however, we will focus on my favorite topic, food! Just a few months ago, the first CRISPR gene-edited food went on the market! In Japan, Sicilian Rouge tomatoes are now being sold after the Tokyo-based company, Sanatech Seed, edited them to contain an increased amount of y-aminobutyric acid (GABA). “GABA is an amino acid and neurotransmitter that blocks impulses between nerve cells in the brain” (https://www.scientificamerican.com/article/crispr-edited-tomatoes-are-supposed-to-help-you-chill-out/). It supposedly (there is scarce scientific evidence of its role as a health supplement) lowers blood pressure and promotes relaxation. In the past, bioengineers have used CRISPR technology to “develop non-browning mushrooms, drought-tolerant soybeans and a host of other creative traits in plants,” but this is the first time the creation is being sold to consumers on the market (https://www.scientificamerican.com/article/crispr-edited-tomatoes-are-supposed-to-help-you-chill-out/)!

Tomatoes

So, how did Sanatech Seed do it? They took the gene editing approach of disabling a gene with the first method described above, making a single cut in the DNA. By doing so, Sanatech’s researchers inactivated the gene that “encodes calmodulin-binding domain (CaMBD)” in order to increase the “activity of the enzyme glutamic acid decarboxylase, which catalyzes the decarboxylation of glutamate to GABA, thus raising levels of the molecule” (https://www.scientificamerican.com/article/crispr-edited-tomatoes-are-supposed-to-help-you-chill-out/). These may seem like big words, but we know from biology that enzymes speed up reactions and decarboxylation is the removal of carbon dioxide from organic acids so you are already familiar with most of the vocabulary! Essentially, bioengineers made a single cut in DNA inside of the GABA shunt (a metabolic pathway) using CRISPR technology. They were therefore able to disable the gene that encodes the protein CaMBD, and by disabling this gene a certain enzyme (glutamic acid decarboxylase) that helps create GABA from glutamate, was stimulated. Thus, more activity of the enzyme that catalyzes the reaction of glutamate to GABA means more GABA! If you are still a little confused, check out this article to read more about how glutamate becomes GABA which will help you better understand this whole process – I know it can be hard to grasp!

After reading all of this research, I am sure you are wondering if you will soon see more CRISPR-edited food come onto the market! The answer is, it depends on where you are asking from! Bioengineered crops are already hard to sell – many countries have regulations against such food and restrictions about what traits can actually be altered in food. Currently, there are some nutritionally enhanced food on the market like soybeans and canola, and many genetically modified organisms (GMOs), but no other genome-edited ones! The US, Brazil, Argentina, and Australia have “repeatedly ruled that genome-edited crops fall outside of its purview” and “Europe has essentially banned genome-edited foods” (https://www.scientificamerican.com/article/crispr-edited-tomatoes-are-supposed-to-help-you-chill-out/). However, if you are in Japan, where the tomatoes are currently being sold, expect to see many more genome edited foods! I know I am now hoping to take a trip to Japan soon!

Thank you so much for reading! If you have any questions, please ask them below!

Embryo Gene Editing can Ensure Offspring Do Not Inherit a Deafness Gene!

Denis Rebrikov, A scientist in Russia has done research regarding ways in which he can edit the genome sequence of an embryo in order to prevent the fetus from developing certain gene mutations, specifically in this case a hearing problem or possible complete deafness. His plans are very controversial to some, who believe the possible risks of very harmful mutations to DNA that would be passed onto direct and future offspring, outweigh the possible benefits. However, some people find this scientific possibility to be worth the risk, if it means not passing a potentially very harmful gene down to offspring. If these methods are done correctly, it should alter the genome sequence in the embryo so that future offspring off that embryo will not inherit the negative mutation.

One couple shared their story in detail, in which both parties have a hearing deficiency, the man with partial deafness, and the woman completely deaf. Their biggest hope is to have children who will not inherit hearing issues, because of the apparent challenges they have had to face themselves because of them. They would be the first couple to perform this gene editing on an IVF embryo, so they obviously have some reservations. One of those being publicity, but more importantly the potential risks of using the CRISPR genome editor. They already have a daughter with hearing loss, but they never chose to test her genes for mutations, nor did they get her a cochlear implant to aid her hearing, because of the potential risks of that. When they finally tested her genes, they learned that she had the same common hearing loss mutation called 35delG in both her copies of a gene called GJB2. The parents then tested themselves, realizing they were both 35delG homozygous, meaning their daughter’s mutations were not unique to her, they had been inherited.

If either the mother or father had a normal copy of the GJB2 gene, a fertility clinic could have more easily created embryos by IVF and tested a few cells in each one to select a heterozygote–with normal hearing–to implant. At this stage, Denis Rebrikov informed them that CRISPR genome editing would be their only option. However, the process presents possibly deal breaking risks, such as mosaicism, in which a gene edit might fail to fix the deafness mutation, which could create other possible dangerous mutations like genetic disorders or cancer. The couple has not decided to go through with the editing just yet, but it is something they are open to in the future as more possible new research or test subjects become available.

Explaining the CRISPR Method: “The CRISPR-Cas9 system works similarly in the lab. Researchers create a small piece of RNA with a short “guide” sequence that attaches (binds) to a specific target sequence of DNA in a genome. The RNA also binds to the Cas9 enzyme. The modified RNA is used to recognize the DNA sequence, and the Cas9 enzyme cuts the DNA at the targeted location… Once the DNA is cut, researchers use the cell’s own DNA repair machinery to add or delete pieces of genetic material, or to make changes to the DNA by replacing an existing segment with a customized DNA sequence.” -US National Library of Medicine Genetics Home Reference

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Woman with a hearing aid 

If you had the opportunity to alter something in the gene’s of your baby’s embryo, would you? Under what circumstances would you consider this, and what risks might stop you from deciding to do it? Comment down below.

 

 

New anti-CRISPR Proteins Serving as Impediments to this Miraculous System.

CRISPR-Cas9 systems are bacterial immune systems that specifically target genomic sequences that in turn can enable the bacterium to fight off infecting phages. CRISPR stands for “clusters of regularly interspaced short palindromic repeats” and was  first demonstrated experimentally by Rodolphe Barrangou and a team of researchers at Danisco. Cas9 is a protein enzyme that is capable of cutting strands of DNA, associated with the specialized stretches of CRISPR DNA.

Diagram of the CRISPR prokaryotic antiviral defense mechanism.

Recently, a blockage to the systems was found by researchers which are essentially anti-CRISPR proteins. Before, research on these proteins had only showed that they can be used to reduce errors in certain genome editing. But now, according to Ruben Vazquez Uribe, Postdoc at the Novo Nordisk Foundation Center for Biosustainability (DTU), “We used a different approach that focused on anti-CRISPR functional activity rather than DNA sequence similarity. This approach enabled us to find anti-CRISPRs in bacteria that can’t necessarily be cultured or infected with phages. And the results are really exciting.” These genes were able to be discovered by DNA from four human faecal samples, two soil samples, one cow faecal sample and one pig faecal sample into a bacterial sample. In doing so, cells with anti-CRISPR genes would become resistant to an antibiotic while those without it would simply die. Further studies found 11 DNA fragments that stood against Cas9 and through this, researchers were ultimately able to identify 4 new anti-CRIPRS that “are present in bacteria found in multiple environments, for instance in bacteria living in insects’ gut, seawater and food,”  with each having different traits and properties.  “Today, most researchers using CRISPR-Cas9 have difficulties controlling the system and off-target activity. Therefore, anti-CRISPR systems are very important, because you want to be able to turn your system on and off to test the activity. Therefore, these new proteins could become very useful,” says Morten Sommer, Scientific Director and Professor at the Novo Nordisk Foundation Center for Biosustainability (DTU). Only time will tell what new, cool, and exciting discoveries will be made concerning this groundbreaking system! What else have you guys heard? Comment below!

Let’s Talk About Malaria

Let’s Talk About Malaria

A small mosquito landing on a human finger.

 

Did you know, that the World Health Organization estimates that roughly 438,000 people die annually due to Malaria? Well, now you do know that unfortunate fact. But – did you know that the total number of people affected by malaria is only growing? In reality, those don’t matter, what does matter is what we are going to do now to combat the issue and CRISPR/cas9 might be the answer. In order to better understand the issue of Malaria and the resolution of utilizing CRISPR/cas9, let’s take an indepth look at both with the assistance of the article about Gene Editing to end Malaria from Vox.

 

So, what is Malaria? According to the Center for Disease Control, Malaria is a mosquito-borne disease caused by a parasite. The four kinds of malaria parasites that infect humans are Plasmodium falciparum, P. vivax, P. ovale, and P. malariae. Typical symptoms causes people to experience fever, chills, and flu-like illness. Left untreated, they may develop severe complications and die. Basically, Malaria has been affecting the global population for decades.  Now, you might be asking yourself: then, what is CRISPR/cas9? Fantastic Question! According to the National Institute of Health, CRISPR/cas9 is recent biomedical technology phenomenon that is drastically changing the genome editing space. In specifically, CRISPR/cas9, which is short for clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9, has generated a lot of excitement in the scientific community because it is faster, cheaper, more accurate, and more efficient than other existing genome editing methods.

 

So, here is the big question: why does it matter? Here is why it matters. When looking at Anopheles Gambiae Mosquito larvae, a common carrier of the Malaria parasite, in a lab in the United Kingdom, a couple of researchers noticed that all of the larvae had a physical red fluorescent phenotype. Although this doesn’t sound shocking, this is extremely shocking as only one parent had the red fluorescent recessive genotype and the other had the dominant wild type, so the expected offspring would be fifty percent with the red fluorescent gene and fifty percent without the red fluorescent gene, but all of the Mosquitos had the red fluorescent gene. This gene has been linked on Mosquitos to the fertility of female mosquitos. Now, you might be asking yourself: when does CRISPR/cas9 come into play? Well, CRISPR/cas9 can target and locate a specific gene, cut, enter itself in and then passed onto the abundant and constant offspring. As a result, when the CRISPR/cas9 is utilized to alter the mosquito population to be resilient to the Malaria parasite and could “wipe” Malaria from the future history of the planet.

 

In reality, I could never say that this is bad thing as it is working to save lives of hundreds of thousands of people globally. As a matter of fact, I would believe the majority of the population would say this is a good thing, but I am going to say this: do it, but do it right. This is something that needs to be done, Malaria has wreaked havoc on our global community for decades and we must move past that, but any small mistake would halt progress in this field for year. In conclusion, let’s keep having a serious discussion on changing the status of Malaria globally.

 

Thank you!

 

From your favorite bacteria,

SAMonella

 

Using CRISPR to target neurons

A rat brain stained with protein and DNA.

Researchers from the University of Alabama at Birmingham have successfully used CRISPR to target neurons. With their novel approach, the team led by Jeremy Day was able to manipulate the function of neurons in vivo.

CRISPR, a self-defense system for bacteria against viral invaders, has become a very popular gene editing tool, as it allows researchers to make very targeted changes to an organism’s DNA. Normally using CRISPR-Cas9, the process involves a piece of guide RNA guiding Cas9 to the desired gene where it cuts it, rendering the gene inexpressible.

However, Day’s team used a different CRISPR mechanism, CRISPRa, which increases the expression of the desired gene. For their CRISPRa, they used CRISPR-dCas9, a CRISPR system with a deactivated Cas9, to which they attached transcriptional effectors. This allowed the guide RNA to guide the transcriptional effectors to a particular gene so it could be up-regulated, increasing its expression. In focusing on neurons, Day’s team targeted the promoter sequence for SYN genes, a common group of genes in the brain that code for proteins that regulate neurotransmitters, and designed their guide RNA accordingly.

After injecting their effector-coupled dCas9 system into live rats using viral hosts, the desired genes were successfully up-regulated, with the researchers viewing their new protein products after the fact through fluorescent markers in cell samples. Following this achievement, Day and his team expanded their CRISPR-dCas9 system, incorporating multiple guide RNAs into a single system to target multiple sections of DNA at the same time and using it analyze the complex Bdnf gene that has multiple promoters and plays a core role in brain function and development.

This innovative approach to targeting genes in the brain has far-reaching applications, allowing for versatile gene editing in live animals, which, in the words of Vanderbilt Brain Institute researcher Erin Calipari, “is going to give us an unprecedented view of the role of gene expression in behavior”.

From psychology to physiology and beyond, there is no doubt that this discovery’s molecular insight will give us a far greater understanding of the brain.

CRISPR/Cas9: A New Means to Alter Genes

Biologists can now control genetic inheritance in mammals with a CRISPR/Cas9-based approach, which allows geneticists to alter parts of the genome by removing, adding or altering sections of the DNA sequence.  Scientists have sought a way to make precise changes to the genome of living cells for a long time, and now they actually can. You may ask, what are CRISPR and CAS9? Why are they important? Simply put, “The functions of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and CRISPR-associated (Cas) genes are essential in adaptive immunity in select bacteria and archaea, enabling the organisms to respond to and eliminate invading genetic material.”  Thus, this recent discovery has created the groundwork for developing new ways to fight diseases. UC San Diego researchers are responsible for this breakthrough. First, they injected a mouse with an engineered active genetic “CopyCat” DNA element into a Tyroisinase gene. The Tyroisinase gene determines fur color. The CopyCat element “disrupts” both copies of the Tyroisinase gene, causing the mouse to have white fur instead of black. The CopyCat element, however, could not spread through a population by itself, unlike the CRISPR/Cas9 systems, which could. This approach, though, was effective only in female mice, not in male ones, likely because of timing differences in meiosis – “a process that normally pairs chromosomes to shuffle the genome and may assist this engineered copying event.” The findings are nonetheless a success. Scientists are optimistic they will be able to alter multiple genes and traits using the same techniques in the near future. Cooper, one of the researches, summed up their achievement nicely: “We’ve shown that we can convert one genotype from heterozygous to homozygous. Now we want to see if we can efficiently control the inheritance of three genes in an animal. If this can be implemented for multiple genes at once, it could revolutionize mouse genetics,” said Cooper. More importantly, these findings continue to speed up research into diseases like cancer and mental illness.

Related image

CRISPR-CAS9 — “How the genome editor works”

CRISPR Technology is Finding Its Place in the Agricultural Industry

CRISPR technology is now laying a foundation in the agricultural world, trying to help corn growers improve the speed, versatility, and output of their crops. It has been difficult to implement CRISPR technology thus far, as the cells walls of plants, at a microscopic level, are particularly tough to penetrate. Fundamentally, CRISPR “…consists of enzymatic scissors called Cas9 that a guide made from RNA shuttles to an exact place in a genome.” The difficulty with plants cells is that, in comparison to animal cells, the extra-rigid cell walls make it immensely difficult for the guide RNA (gRNA) and the Cas9 to reach their destination on the genome. In response to this problem researchers have come up with what is described as an “inelegant” solution to this problem where they “…splice […] CRISPR genes into a bacterium that can breach the plant cell wall or put them on gold particles and shoot them with what’s known as a gene gun.” Unfortunately, this method doesn’t work in the crucial corn varieties where it is needed. However, a team of researchers in North Carolina, Timothy Kelliher and Quideng Que of Syngenta, in Durham, North Carolina have come up with an even more ingenious solution to deal with the stubborn plant cell walls. Haploid induction “…allows pollen to fertilize plants without permanently transferring ‘male’ genetic material to offspring. The newly created plants only have a female set of chromosomes – making them haploid instead of the traditional diploid. Haploid induction itself can lead to increased breeding efficiency and higher yielding plants.” This same method has been found to work in wheat and even Arabidopsis, “…a genus of plants related to cabbage, broccoli, kale, and cauliflower.” Yet again, sadly, CRISPR faces another drawback as scientist not that “…if it were done in the field, the changes wouldn’t spread because the male genome in the pollen – which carries the CRISPR apparatus – disappears shortly after fertilization.” However, there is still much hope for CRISPR technology, and it is without a doubt that we are making big strides into the future with gene editing technology.

Fighting the mosquito disease problems with… mosquitos?

Since the discovery of CRISPR-Cas9 system (Clustered Regularly Interspaced Short Palindromic Repeats), gene editing has become a highly debated topic. One of the reasons backing the use of CRISPR-cas9 is to prevent diseases. These diseases include mosquito-borne diseases such as zika, dengue fever, and malaria.  Malaria in particular kills around 3,000 children every year. Various groups of scientists have worked on genetically modifying mosquitos to stop the spread of malaria by making female offspring sterile and unable to bite, making male offspring sterile, or making mosquitos resistant to carrying diseases. A point of concern was if the modified gene would stay relative and would carry from generations. In order to make offspring, genes from both parents must be used, resulting in the offspring carrying the modified gene only half the time.  In particular cases, mutations would occur in the altered DNA, which nullified the genetic changes.  This has been solved by developing a gene drive, which makes the desired gene dominant and occur in the offspring almost 100% of time.  This entails almost the entire mosquito population could have this modified gene in as little as 11 generations.

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Recently, the government of Burkina Faso, a small land-locked nation in west Africa, has approved for scientists to release mosquitos that are genetically modified anytime this year or next year.  The particular group of mosquitos to be released first is a group of sterile males, which would die rather quickly.  Scientists want to test the impact of releasing a genetically modified eukaryotic organism in the Africa. It is the first step in “Target Malaria” project to rid the region of malaria once and for all.

 

One of the major challenges in gaining allowance to release the genetically modified species was the approval of the residences, who lack words in the local language to describe genetics or gene editing.  Lea Pare, who leads a team of scientists modifying mosquitos, is working with linguists to answer questions the locals may have and tp help develop vocabulary to describe this complex scientific process.

What do you think about gene editing to possibly save millions?

Read the original article here.

View a video explaining how scientists can use genetic engineering to fight disease here.

A Treat for the Muscles!

Scientists using CRISPER-Cas9 gene-editing technique have managed to better the lives of four dogs suffering from the most common form of muscular dystrophy, Duchenne.

https://commons.wikimedia.org/wiki/File:Smiling_Dog_Face.jpg

A research team led by U.T Southwestern Medical Center edited muscle cells in young dogs with Duchenne to remove a short, problematic segment of protein-coding DNA that occurs in both canine and human patients. Within about two months, the dogs were producing a greater amount of dystrophin.

To get this gene-editing technology into the dog’s muscles, the research team created viruses to transport the gene-editing machinery. To do so, the scientists had to extract some of the virus’s own DNA in order to fit the gene-editing machines. The viruses were assigned either of two tasks. Some viruses carried Cas9-molecular “scissors” to cut out the DNA sequence that blocks the production of dystrophin in muscle cells. The other viruses carried a guide molecule to help the Cas9 to identify where it should make those cuts.

Using viruses as a means to transport the gene-editing technology is very helpful because viruses are very small, even smaller than bacteria. When a virus enters your body, it invades some of the cells and takes control of the cell’s functions by injecting its genetic materials into the cell.

For now, the research team has already demonstrated that CRISPR can treat Duchenne in human cells in the lab, but this test was the first success with a large mammal. Adding on, for this study, the research team focused on the protein level, not on how this treatment may have affected the dog’s behavior.

To conclude, one question that remains in the air after this demonstration is how long one injection with CRISPR will last in human Duchenne patients versus dogs. The research team is hoping once, but there is still so much to discover!

CRISPR/CAS9: Potential to destroy malaria?

CRISPR. Sounds more like a new brand of potato chip than something potentially revolutionary (Bold new flavor. Bold new crunch. CRISPR.). Nevertheless, this tool used for easy gene editing and slicing is tearing up the science world because it could be the key to combatting disorders and diseases.

Recent research indicates that CRISPR/Cas9 based genome editing tools could aid in the fight against malaria, one of the “big three” diseases that has long affected and continues to affect humans worldwide. How is CRISPR/Cas9 able to do this?

CRISPRs (Clustered Regularly Interspaced Short Palindromic Repeats) originally are how bacteria protect themselves from foreign viruses. CRISPRs contain DNA from viruses that have attacked the bacteria, and so when a similar virus attacks, the bacteria knows that this virus and his DNA are bad. Essentially, CRISPRs allow bacteria to build up immunity. When foreign DNA is detected, the Cas9 enzyme is guided by the CRISPR and is able to cut the desired DNA. Scientists have come up with a way to engineer and manipulate the CRISP/CAS9 system into other organisms (such as mosquitoes) so that we can successfully edit genome sequences and genes to produce desired results. We take advantage by specifying which genes the Cas9 should cut/replace, and then it does just that. Therefore, the CRISPR/Cas9 system allows us new genome editing potential like none before.

Made by Viktoria Anselm.

How does this apply to mosquitoes and malaria? Scientists experimented with genetically modified malaria-transmitting mosquitoes (Anopheles gambiae), altering the fibrinogen-related protein 1 (FREP1) gene on them. This gene essentially codes for a protein that makes mosquitoes a vector for malaria. The scientists used the CRISPR/Cas9 to inactivate this gene.

The results produced mosquitoes with significantly less transmission of malaria to both human and rodent cells. However, these mosquitoes have “reduced fitness”: a significantly lower blood-feeding propensity, egg hatching rate, a retarded larval development, and reduced longevity after a blood meal. Essentially this means that these mosquitoes have a low chance of affecting populations of mosquitoes in the wild without being “pushed” by scientists, where scientists are “forcing DNA to inherit particular sets of genes.” This is called a gene drive. With a strong push for a couple of years, there is potential for worldwide mosquito populations to be significantly changed in 10-15 years.

Photo taken by JJ Harrison

I chose to write about this new research and potential breakthrough because it really means something to me, as I have lived in and visited countries threatened by malaria. I had to take preventative pills every morning, and I would have to sleep in a restrictive mosquito net. All that made me wonder about and feel for a kid in the same country who didn’t have those things and how he or she would manage without those barriers to malaria. Having said that, I really do believe this is a worthwhile option we should explore, and I think it can make a difference for the world.

What do you think? Do you think it is realistic for theses mosquitoes to change the entire mosquito population and effectively help reduce malaria transmission? Will CRISPR/Cas9 work as we hoped? Or is it too good to be true?

Cas9: A Clue Into Making Gene Editing Safer

CRISPR is a revolutionary system that edits the DNA of living organisms with ease. The gene-editing technology offers scientists insight into genetic diseases and is widely used in biotech and agriculture, as well as to treat cancer and viral infections. But the CRISPR system and its mechanisms are not yet fully understood. However, researchers at the Ohio State University have reported that they have figured out the mechanism of how the CRISPR system figures out where and when to cut the DNA strands. This is particularly revolutionary as it provides insight into preventing gene-cutting errors.

Cas9 is an enzyme that is used by the system to target and cut out or insert specific genes. In the second of two paper published in the Journal of the American Chemical Society, the team invalidates the widely-held belief that the enzyme cuts DNA evenly. Professor of chemistry and biochemistry Zucai Suo explains that instead of cutting both sides of the DNA double-helix to the same length, Cas9  actually trims each side to uneven lengths. Ohio State doctoral student Austin Raper and his co-authors determined that the two different parts of the “Cas9 molecule communicate with each other to set the location and timing of a cut”. The first part of the molecule sets forth to cut its respective DNA strand and changes shape and signals to the second part to cut its respective second strand.

Crystal Structure of Cas9 Enzyme

Suo says that he hopes their work allows for scientists to minimize and eventually eliminate gene-editing errors. CRISPR rarely target unintended genes, but gene-editing errors can have very serious consequences. For example, if the system accidentally cut a tumor suppressor gene from a person’s DNA, they would be much more likely to develop cancer. As Raper says, it is important to understand CRISPR and the Cas9 enzyme mechanisms in order to allow CRISPR to advance to its full potential.

 

Source: https://news.osu.edu/news/2018/02/28/cas9-2cuts/

Deleting Genes to Stop Malaria

A new discovery has highlighted the positive effects that the revolutionary new gene editing tool, CRISPR-Cas9, can have. Scientists at the Johns Hopkins Bloomberg School of Public Health’s Malaria Research Institute have discovered that the deletion of a single gene from the Anopheles Gambiae mosquito, called the FREP1 gene, yields promising results in the eradication of the malaria disease.

 

Image result for mosquito gene editing

Gene Editing

The FREP1 gene has been associated with being a malaria “host factor” gene because it helps the parasite live in the gut of the mosquitoes.  However, the scientists, using the CRISPR-Cas9 gene editing procedures, have been able to delete the FREP1 gene from the mosquitoes and have seen significant decreases in the spread of malaria. Without the host factor gene, the parasite has difficulty surviving in the mosquito, which decreases the spread of the disease to other organisms.

 

The deletion of the FREP1 gene had other effects in addition to the resistance of the malaria parasites. In the mosquitoes where the gene was deleted, many showed no signs of sporozoite-stage parasites in their salivary glands, which can spread to humans through mosquito bites. George Dimopoulos, PhD, professor in the Bloomberg School’s Department of Molecular Microbiology and Immunology, commented on the study, saying that “if you could successfully replace ordinary, wild-type mosquitoes with these modified mosquitoes, it’s likely that there would be a significant impact on malaria transmission”.

The Future of CRISPR

CRISPR is starting to become more and more of a reality as Harvard professor David Liu continues to work on it. Liu was the person who originally developed CRISPR first base editor which allowed for single letter changes in the genetic code. Liu has come up with two new features to CRISPR-Cas9.

The first is called cellular detective or CAMERA(CRISPR-mediated analog multievent recording apparatus systems). What this function does is it finds the genetic problem that is responsible for the disease someone is experiencing. Cas9 will record all the cell data and piece info together, which overall will provide more information about cancer, stem cells, aging, and overall disease.

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The second finding is referred to as sharp scissors which is a CRISPR enzyme. Sharp scissors are way more precise and accurate than the old enzyme making is much safer. The scissors depend on specific DNA to find the region where it is supposed to cut or edit. CRISPR is progressing and as more research is being done could be used on humans in the future.

 

New Enzyme Reducing Off-Target DNA Editing

A new enzyme named xCas9 allows researchers to target more sites in the genome than with traditional CRISPR-Cas9 editing, while also reducing off-target effects. The technique was reported earlier this year (February 28) by a biologist David Liu and his colleagues.

CRISPR-Cas9 has become the gene-editing tool of choice in many labs due to the effectiveness and convenience. But CRISPR-Cas9 has limitations like the necessity of targeting a particular sequence called a PAM near the gene to be modified, which limits researchers’ ability to make specific genetic changes.

“Relief from the PAM restriction is quite important,” Albert Jeltsch of the University of Stuttgart in Germany. “Some of these elements are quite small, and then the restriction can be quite relevant.”

Liu and his colleagues used a laboratory technique to evolve an enzyme that could recognize a broader range of PAM sequences, enabling more sites in the genome to be targeted. It just so happens that xCas9 also turned out to be more specific to the targeted sites, with fewer off-target effects. xCas9 will allow gene therapy to have higher success rates.

The Miracle of CRISPR/Cas9 in Gene Editing

Some scientists say, “you can do anything with CRISPR” and others are absolutely astonished and amazed.

CRISPR can rapidly change any gene in any animal or plant with ease. It can fix genetic diseases, fight viruses, sterilize mosquitos and prepare organs for transplant. The possibilities are endless – and the prospect of designer babies isn’t far off.

https://en.wikipedia.org/wiki/CRISPR#/media/File:Crispr.png

Dead Cas9 can fix a single base pair typo in DNA’s genetic instructions. It can convert a C-G into a T-A pair. Also, we can attach fluorescent tags to dead Cas9 so researchers can locate and observe DNA or RNA in a living cell. Dead Cas9 can also block RNA Polymerase from turning on a gene, in CRISPRi. In CRISPRa, a protein that turns on genes is fused to dead Cas9.

CRISPR can be used for anything involving cutting DNA. It guides molecular scissors (Cas9 enzyme) to a target section of DNA & works to disable or repair a gene, or insert something new.

Many scientists have been thinking of improvements for this miracle gene editor. RNA Biologist Gene Yeo compares the original Cas9 to a Swiss army knife with only one application – a knife. He says that by bolting other proteins and chemicals to the blade, they transformed the knife into a multifunctional tools.

CRISPR/Cas9 is special because of its precision. It is much easier to manipulate and use compared to other enzymes that cut DNA. By using “guide RNA” it can home in on any place selected by the researcher by chemically pairing with DNA bases.

While Cas9 does have some problems, scientists definitely see the potential for greatness with a few tweaks. They wanted to ensure permanent single base pair changes, and they increased that from 15 to 75 percent. Liu used a hitchhiking enzyme called cytidine deaminase.

Scientists researched chemical tags on DNA called epigenetic marks. When scientists placed the epigenetic marks on some genes, activity shot up. This provided evidence that the mark boosts gene activity.

Case can also revolutionize RNA biology. The homing ability of CRISPR/Cas9 is what makes this seem possible. It was found that Cas9 could latch on to mRNA.

CRISPR/Cas9 was first found in bacteria as a basic immune system for fighting viruses. It zeroes in on and shreds the viral DNA. Half of bacteria have CRISPR immune systems, using enzymes beyond Cas9.

Overall scientists predict that in the next few years, results will be amazing. The many ways of using CRISPR will continue to multiply and we will see where science takes us.

Source: https://www.sciencenews.org/article/crispr-inspires-new-tricks-edit-genes

Other Sources: https://www.neb.com/tools-and-resources/feature-articles/crispr-cas9-and-targeted-genome-editing-a-new-era-in-molecular-biology

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