Gene-editing – BioQuakes

Tag: Gene-editing

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

On March 17, 2019

In Student Post

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

On March 12, 2019

In Student Post

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

On March 12, 2019

In Student Post

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

By vonstadenine

On March 12, 2019

In Student Post

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.

CRISPR-CAS9 — “How the genome editor works”

CRISPR Technology is Finding Its Place in the Agricultural Industry

By xraychristollography

On March 12, 2019

In Student Post

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?

On October 10, 2018

In Student Post

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.

Image by Author

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!

By frantibiotic

On October 10, 2018

In Student Post

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?

On March 12, 2018

In Student Post

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

By sharothymine

On March 12, 2018

In Student Post

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

On March 12, 2018

In Student Post

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

On March 11, 2018

In Student Post

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.

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

On March 10, 2018

In Student Post

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

On April 5, 2017

In Student Post

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

Anti-CRISPR Proteins: What are they and can they be beneficial?

On April 5, 2017

In Student Post

NIH Image Gallery Image Link

Understanding CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)

For many bacteria, one line of defense against viral infection is the RNA guided “immune system” known as CRISPR-Cas. This particular complex is unique because of its ability to recognize viral DNA and trigger its destruction. Scientists have used CRISPR to degrade sections of viral RNA and use the CRISPR systems to remove unwanted genes from an organism. CRISPR proteins have also been studied with the hope of eliminating serious disease and illnesses. However, this CRISPR system does not always work do to anti-CRSPR proteins that inhibit the complex from working properly.

Research

According to an article on ScienceDaily, researchers have finally discovered how these anti-CRISPR proteins work! Research done by biologist Gabriel C. Lander from the Scripps Research Institute, discovered that anti-CRISPR proteins work by inhibiting CRISPR’s ability to identify and attack viral genomes. Just like there are different CRISPR systems, there are multiple anti-CRISPR proteins as well. One in particular mimics DNA to throw the CRISPR-guided detection machine off its course. Scientists have been able to further discover certain aspects of CRISPR and anti-CRISPR systems by using a high-resolution imaging technique called cryo-electron microscopy. They have discovered that the CRISPR surveillance complex analyzes a virus’s genetic material to see where it should attack by having proteins within the complex wrap around the CRISPR RNA, exposing specific sections of bacterial RNA. These sections of RNA then scan viral DNA, looking for genetic sequences they recognize. Lander describes these proteins as being very clever because they “have evolved to target a crucial piece of the CRISPR machinery. If bacteria were to mutate this machinery to avoid viral attacks, the CRISPR system would cease to function.” Therefore, CRISPR systems cannot avoid anti-CRISPR proteins without completely chancing the mechanism used to recognize DNA. Another type anti-CRISPR protein works a bit differently. Based on its location and negative charge, this anti-CRISPR protein acts as a DNA mimic, fooling CRISPR into binding this immobilizing protein, rather than an invading viral DNA.

Can Anti-CRISPR Proteins be beneficial?

Researchers are saying that the understanding of how these anti-CRISPR proteins work are extremely important! According to an article on GEN, the discovery and understanding of anti-CRISPR proteins actually allows researchers to have greater control over gene-edits. In this article, Dr. Sontheimer, a professor in the RNA The RNA Therapeutics Institute at UMass Medical School, expressed how “CRISPR/Cas 9 is a good thing because it introduces specific chromosome breaks that can be exploited to create genome edits, but because chromosome breakage can be hazardous, it is possible to have too much of a good thing, or to have it go on for too long.” Anti-CRISPR proteins can be beneficial and work as an off switch for CRISPR, therefore advancing gene editing!

The Grey Area of Human Gene Editing

By cottoneaster

On March 1, 2017

In Student Post

The process of Human Gene Editing developed with the goal to prevent future generations from suffering from genetic diseases present in past generations, like our own. Human gene editing, provided it is done only to the correct disease, alters the DNA in embryos, eggs, and sperm to the when reproduction occurs, the gene for the disease or disability is not inherited. However, two weeks ago the National Academies of Sciences and Medicine issued a report stating that human gene editing is being used to enhance people’s health or abilities. This is considered unethical according to organizers of a Global Summit on human gene editing.

Human gene editing has been given a “yellow light” because the process is not yet approved to be done on people. There are high hopes that diseases caused by only 1 genetic mutation such cystic fibrosis and Huntington’s disease will be eliminated due to this process. Unfortunately diseases that are caused by more than one genetic mutation, such as autism or schizophrenia, are not curable by this process.

National Cancer Institute

Gene Editing on humans is such a controversial topic right now: is it ethical to change genes? should the practice be used to change physical appearances? Ultimately, if Human Gene Editing is approves, who decides when it becomes too much, or unethical. This grey area is presented to be somewhere between when it is appropriate to help aid the life of a human, ridding them of a disease, and when enhancements are made.

More CRISPR Improvements

In Student Post

Crispr-Cas9 is a genome editing tool that is creating a whole lot of buzz in the science world. It is the newest faster, cheaper and more accurate way of editing DNA. Crispr- Cas9 also has a wide range of potential applications. It is a unique technology that enables geneticists and medical researchers to edit parts of the genome by cutting out, replacing or adding parts to the DNA sequence. The CRISPR-Cas9 system consists of two key molecules that introduce a mutation into the DNA. The first Molecule is an enzyme called Cas9. Cas9 acts as a pair of scissors that can cut the two strands of DNA at a specific location in the genome so that bits of DNA can be added or removed. The second is a piece of RNA called guide RNA or gRNA. This consists of a small piece of pre-designed RNA sequence located within a longer RNA scaffold. The scaffold part binds to DNA and the pre-designed sequence guides Cas9 to the right part of the genome. This makes sure that the Cas9 enzyme cuts at the right point in the genome.

CRISPR-Cas9 is efficient compared to previous gene-editing techniques, but there’s still plenty of room for improvement. CRISPR is less efficient when employing the cellular process of homology-directed DNA repair, or HDR, as opposed to nonhomologous end joining. Jacob Corn, the scientific director of the Innovative Genomics Initiative at the University of California, Berkeley, and his colleagues have come up with a way to improve the success rate of homology-directed repair following CRISPR-Cas9. “We have found that Cas9-mediated HDR frequencies can be increased by rationally designing the orientation, polarity and length of the donor ssDNA to match the properties of the Cas9-DNA complex,” the researchers wrote in their paper, “We also found that these donor designs, when paired with tiled catalytically inactive dCas9 molecules, can stimulate HDR to approximately 1%, almost 50-fold greater than donor alone.”

“Our data indicate that Cas9 breaks could be different at a molecular level from breaks generated by other targeted nucleases, such as TALENS and zinc-finger nucleases, which suggests that strategies like the ones we are using can give you more efficient repair of Cas9 breaks,” coauthor Christopher Richardson, a postdoc in Corn’s lab, said in a statement.

Original Article:

http://www.the-scientist.com/?articles.view/articleNo/45159/title/More-CRISPR-Improvements/

Other Addtional Helpful Links:

http://www.yourgenome.org/facts/what-is-crispr-cas9

Should We Use It: Crispr-Cas 9 Edition

By mosmosisjones

On April 13, 2016

In Student Post

https://www.youtube.com/watch?v=7C20s4eAGtU

Arguably the greatest thing to happen to genetics since the Human Genome Project, Crispr-Cas 9 has been getting a lot of attention. The Los Angeles Times wrote an article approximately 4 months ago discussing the ins and outs of the new gene editing breakthrough. The concept of editing genes is nothing new for scientists. They’ve been doing it since the 1970s. So many people are asking “What makes Crispr so special?” The answer is convenience. Crispr-Cas 9, although still filled with flaws, is the easiest gene editing tool to use out there right now. Scientists from UC Irvine and UC San Diego have used it on mosquitoes to fight malaria and scientists have begun to use it on human embryos as well. Crispr is an acronym for Clustered Regularly Interspaced Short Palindromic Repeats which is a relatively complicated way of saying “gene editing tool.” What Crispr does is it can target certain parts of a strand of DNA and “delete” them from the strand. In reality they aren’t being “deleted” but “turned off” so RNA doesn’t code it and begin to manufacture proteins for it. But the real question is why certain people are against gene editing. Everyone’s seen the movie GATTACA where gene editing is not only commonplace, but discriminatory. However in today’s world, the fear is much more strongly rooted than a fear of “geneticism” (genetic rascism). Using Crispr on viable human embryos to edit genes may have undesired effects. The turning off/on of one gene could result in the unintentional turning on/off of another. Also, many scientists believe that a parent making decisions for an unborn child can be unethical and unfair if the child did not want those changes to be made. And who knows, maybe in the future with the continuous use of Crispr and the development of more complex gene editing tools, “geneticism” could be a reality.

Other articles pertaining to Crispr are linked here and here for more information on the subject.

The New and Improved CRISPR-Cas9

On April 13, 2016

In Student Post

The CRISPR-Cas9 genome editing system has transformed into an even better version of itself. A new, elegant technique, coined by researches at the Broad Institute of MIT and Harvard and the McGovern Institute for Brain Research at MIT, has resolved one of the most reoccurring technical issues in genome editing.

https://commons.wikimedia.org/wiki/File:Crystal_Structure_of_Cas9_in_Complex_with_Guide_RNA_and_Target_DNA.jpg

Primarily, the CRISPR-Cas9 system works to specifically modify a cell’s DNA. CRISPR is dependent on protein Cas9, as it is specialized for cutting DNA. The DNA, at a location identified by a RNA’s sequence matching the target site, is altered by Cas9. Though it very efficient at cutting its target sites, there is a large complication in the process. Once the Cas9 is inside the cell, it can also bind and cut additional sites that are not targeted. Because of this, undesired edits are produced which can alter gene expression or kill off a gene completely. These setbacks can lead to cancer or other problems. Feng Zhang, along with his colleagues at MIT, reported that by just changing 3 out of the approximately 1,400 amino acids composing the Cas9 enzyme from S. pyogenes, a considerable reduction of “off-target editing” to undetectable levels are observed.

This newfound information was derived from studying the structure of the Cas9 protein. Since DNA is negatively charged, it binds to a positively charged groove in the Cas9 protein. The scientists predicted that by replacing some of the positively charged amino acids with a neutral charge, there would be a decrease in binding to “off target” sequences than to “on target” sequences. By mutating three amino acids, their technique proved to be successful.

The team is calling this newly-engineered enzyme “enhanced S. pyogenes Cas9” or “eSpCas9.” It’ll be particularly useful for genome editing that requires precise specificity and it is said to be available for researches worldwide.

I believe that this newfound resolution for the CRISPR-Cas9 genome editing hurtle is a huge game changer. This charge-changing approach might also be able to be used for other experiments involving RNA-guided DNA targeting enzymes. Ethical and societal concerns have also risen due to the idea of rapid and efficient genome editing. The eSpCas9 is highly beneficial in the scientific community, however there is a lot more research needed to be done in order to be used clinically.

Original article can be found here.

CRISPR: Is Science Going Too Far?

On April 13, 2016

In Student Post

CRISPR is a some-what new genetic tool in the field of science to edit human embryos. Using CRISPR, scientists can edit the genes of organisms more precisely than ever before. It uses RNA and an enzyme that slices up invading virusesF. One use of this new technology is to fix mutations that cause genetic diseases.

Crispr

https://en.wikipedia.org/wiki/CRISPR

Ethical concerns arose in April of 2015 when Chinese research used CRISPR to edit nonviable human embryos. In addition, some fear that the use of CRISPR to give the embryo traits not found in their genetic code can lead to a obsessive gene culture like the one found in Gattaca. This ethical debates caused scientists to meet at an international summit hosted by the United States National Academies of Sciences and Medicines, where the scientists discussed the ethical concerns of CRISPR but agreed to continue researching it cautiously.

In addition, some argue that using CRISPR for gene editing defeats the sacredness of the human genome and is unnatural. To this point, Sarah Chan from the EuroStemCell argues, “There is nothing sacred or sacrosanct about the genome as such. The human genome – the genome of humanity as a whole, and the unique individual genome we each possess – is merely the product of our evolutionary history to date”. From this point of view, the genome is merely a record of one’s history, but to some religious groups it is a symbol of life which should not be tainted with.

So readers, what do you think? Should we use this tool to help cure treatable diseases, or does this new technology cross the line between scientific mechanisms and morality? What type of genes should this new tool be allowed to edit?

Other sources

https://www.sciencenews.org/article/year-review-breakthrough-gene-editor-sparks-ethics-debate

http://www.sciencemag.org/news/2016/04/crispr-debate-fueled-publication-second-human-embryo-editing-paper

http://www.wired.com/2015/12/stop-dancing-around-real-ethical-problem-crispr/

http://www.eurostemcell.org/commentanalysis/ethics-changing-genes-embryo

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