Questions tagged [crispr]

Cluster Regularly Interspaced Short Palindromic Repeats; Naturally a prokaryotic adaptive immune system, elements of CRISPR are increasingly being used for sequence specific DNA targeting in biotechnological applications, especially genome editing. This tag can be used for questions about its natural function or biotechnological applications, including elements of the CRISPR system such as Cas9 and Cpf1 nucleases.

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How to design a permanent vaccine to cholera using CRISPR?

I know that Vibrio Cholerae infects the body through the GM1 ganglioside. So, would it be possible to engineer a CRISPR gene editing tool to prevent Vibrio Cholerae from getting into our cells? ...
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How to find the enhancer region of a specific gene?

I am new to these concepts in biology and need some help understanding. My main concern is how to find the enhancer of my gene of interest, specifically the sequence of the enhancer. I am working on a ...
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what's the difference between traditional genetic engineering and and CRISPR?

While at school, I learnt about restriction enzymes and how you could insert foreign DNA into an organism. When I first heard about CRISPR I thought it was very similar except much more precise/...
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How I design gRNA for CRISPR cas3?

I am working on CRISPR cas3 project to knockout virulent genes for that I have to design guide RNA but there is no software available for cas3. how can I design guide RNA?
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Can Cas13 be used with multiple crRNAs in the same reaction?

CRISPR-Cas13 equipped with crRNA (complementary to transcripts of interest) can be designed to target ssRNA transcripts in cells. Upon successful crRNA and ssRNA binding, a fluorescent domain on ...
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CRISPR/Cas9 edited E.coli on AFM?

I am doing CRISPR/Cas9 experiment on E. coli. I am introducing recombinant plasmid BPK764 (which carries Cas9 + sgRNA designed and added later in that plasmid) into compentent E.coli cells which ...
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What is meant by “heads” and “tails” in the context of gene orientation?

I have a hard time understanding what this paper is talking about when it says: We observed maximal cleavage at sites oriented tail-to-tail and separated by -10 bp to +30 bp (Fig. 2d). Finally, ...
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Can CRISPR also remove DNA viruses?

If I'm not mistaken only RNA viruses insert themselves into the host genome. As an example of DNA viruses, herpes viruses for example do not insert themselves in the host genome. Can CRISPR cut DNA ...
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How do DNA viruses keep themselves in the nucleus without inserting themselves into genome?

If I'm not mistaken only RNA viruses insert themselves into the host genome. As an example of DNA viruses, herpes viruses for example do not insert themselves in the host genome. Then how do DNA ...
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DIYbio - CRISPR injection sites for targeting the ABCC11 gene [closed]

I've been researching into the biohacking world where people most notable Josiah Zayner and Tristan Roberts have used a CRISPR solution developed in their backyard for gene therapy. There is even a ...
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Size constraints on CRISPR guide RNA

I had a quick questions on the size limitations of a CRISPR guide. More specifically on the shorter end. Can I make a guide that is say 7-10bp and still have an active complex? I transfect using an ...
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How does a CRISPR therapy work?

Afaik you can use CRISPR to edit genes of a cell. But how should therapy for a human work based on CRISPR? Can a drug perform it on vast numbers of ill cells?
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Translocation and Isolation of Specific Bacterial Codons

I am currently working on a research project in which I am aiming to translocate a gene from a specific bacteria (strain NT-26, an arsenic oxidizing bacterium) into another bacterium. I am doing this ...
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When gene editing are both chromosomes in a pair changed?

Sorry for the possibly confused question, my knowledge of genetics is limited to medical training only but I have a question. Are gene editing techniques such as CRISPR used on both of the ...
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Can CRISPR-cas9 be used to insert a large gene?

I would like to insert a 1500 bp or longer gene to the broken site in E.coli (or may be bacillus subtilis which support NHEJ) after cut with Crispr-Cas9. Is this possible for both NHEJ or HDR?
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Realistic Application of CRISPR in Human Disorders

Human trials recently began to use the genome editing technology CRISPR to treat sickle cell anemia using edited stem cells. Sickle cell anemia is caused by a single DNA Mutation, and is also a ...
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Crispr complex in human cells?

Is the crispr (where the parts of Virus DNA is saved) section of the DNA existing in human cells aswell or is it just in bacteria cells?
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What is the role of CRISPR-dCas9 in gRNA-dCas9 transcription regulator complexes?

In this paper1, I read that mutant versions of Cas proteins such as a deactivated Cas9 (dCas9) are used alongside a guide-RNA (gRNA) to form variants of CRISPR tool that can function as transcription ...
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Are Restriction Enzymes obsolete with CRISPR?

Are Restriction Enzymes obsolete with CRISPR?
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Mathematical models of gene editing using CRISPR

One confusing thing I have found when reading articles about possible CRISPR based gene therapy treatments in humans is that there is vey little discussion about what percentage of your cells will ...
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Does gen editing have any technical cross-species limits?

A surviving GMO put aside, how far could you go in cross-species gen editing? Is it possible for example to introduce plant genes to human DNA and vice versa? Question: from where is gen editing ...
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How was Crisper Cas9 function discovered?

What paper outlines the discovery of how the CRIPER DNA, CrRNA, and Cas9 interact?
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Crispr/CAS9 genome editing is actually processed at which phase of Cell cycle?

all, Is it only happens in certain phases, like S, G1,...or it can happen any time....or maybe, it has some perferences. Put it in another word, does it going to processe edit if the cell is not ...
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How could a species be engineered to go extinct?

Non-biology background here. I read this very interesting article: https://www.wired.com/story/crispr-eradicate-invasive-species/ However I am having a hard time wrapping my head around something: ...
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A reliable source of Cas9?

I live in Egypt, and I do want to perform CRISPR-Cas9 genome modification, but there are no reliable sources of Cas9. I can order some Cas9 online from the US, but not much. So... my question is, is ...
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Can a gene be inactivated using CRISPR if it is not in the interspace of short palindromic repeats?

I have recently studied how CRISPR works but there is something that I do not understand at all. I have heard a lot of people claiming that with this method it is possible to modify any genome by ...
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Is it possible to use virus for genetic modification of embryos during the fetus stage

I know this probably sounds rather hypothetical and not very feasible but I would very like an answer telling why it is possible or not possible and why. With the advancement of crispr and other dna ...
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What impact do genetic engineering techniques have on seed breeders?

In research of seed breeding, I'm trying to understand the impact of genetic engineering techniques like CRISPR (This is the main one as I understand) on traditional seed breeders. Through searching ...
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295 views

Can CRISPR-Cas9 make changes on a living organism?

Can CRISPR-Cas9 make changes on a living organism? What's the limit here? E.g. reversing/restoring hearing loss in a living adult mice.
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How is the Guide RNA created for Crispr?

It seems that to modify DNA a guide RNA and Cripr are introduced. But I'm unable to understand how the Guide RNA is made or created. Is it a simple method which can be done with simple lab equipment ...
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Are exosomes useful as a transfection or delivery mechanism in gene editing?

The use of viruses as transfection or delivery agents for gene editing (CRISPR/cas9, etc) is well known. However, one problem with using viruses to deliver DNA into cells is the possibility of ...
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Where does tracrRNA comes from? [closed]

I'm talking about CRISPR system. I know the crRNA is transcribed from the palindromic repeat and the "spacer" but I don't know where the tracrRNA comes from.
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What does metal-dependent mean?

I was reading about Cas1 and Cas2 and came across this excerpt: ...Cas2 was identified as a metal-dependent endoribonuclease that cleaves ssRNA or dsDNA... What does metal-dependent mean in this ...
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Mutated cell proliferation

Reading Jennifer Doudna's fascinating book on CRISPR. So she describes rare cases where a mutation in a single cell removes the gene responsible for a genetic disease. The cell proliferates and the ...
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Inheriting Modified DNA after CRISPR editing

If CRISPR is used to modify the DNA sequence to cure a disease - say MS in a woman - will the RNA guides also modify the sequence in her eggs so her children could be born without MS inherited from ...
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What does it mean to “write an image and GIF into the DNA of bacteria”?

BBC News recently published an article saying that: An image and short film has been encoded in DNA, using the units of inheritance as a medium for storing information ... The team sequenced the ...
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CRISPR guide RNA design and primer synthesis

I am trying to knockout huntingtin gene from HeLa cells (human epithelial cells). I used CRISPR explorer and benchling.com to determine the best guide RNA sequence that would bind to the target DNA ...
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Webtool to design guide RNA (gRNA) for use with CRISPR-AsCpf1?

My goals are to use a free webtool to: Identify guide RNAs (direct-repeat sequence followed by the targeting sequence) appropriate for use with AsCpf1 in order to target a specific segment of genomic ...
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Could we eradicate mosquitoes? [closed]

Researchers have proposed the application of CRISPR/Cas9 and gene drive to genetically alter wild mosquito populations such that they don't transmit malaria. The government of New Zealand has ...
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Enhanced Spectrum Chloroplasts

As far as I'm aware, chloroplasts produce energy through the use of photosynthesis which essentiall converts CO2 and Water into Glucose (I'm an engineer by trade so really wracking my brains to the ...
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Functions of tracrRNA and crRNA in the CRISPR/Cas9 system

I need some help about Crispr / Cas9. The CRISPR/Cas9 technic consists (for bacterias) in "cut" bacteriophage's DNA, in order to make it unfunctional. When the RNA is transcribed (RNA which is ...
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CRISPR/Cas9: What are the main differences between sgRNA and the Cr:TracrRNA ?

So from what I understand, in gene editing, the CRISPR vector expresses a small RNA sequence comprised of a small guide-RNA that is complementary to your target sequence. The sgRNA comprises a 20 Bp ...
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Why doesn't CAS9 cleave the original CRISPR sequence in the bacterial genome?

CAS9 is the RNA-guided endonuclease that cleaves DNA as specified by the RNA sequence and is used to target viruses that infect bacteria. So why doesn't this RNA+endonuclease combo also cleave the ...
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What is the role of tracrRNA in CRISPR-cas9?

From what I understand, in a CRISPR cas9 complex, gRNA is comprised of tracrRNA and crRNA. I've read that crRNA is the part which is matched to the DNA which is targeted, but what role does tracrRNA ...
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Is Cas9 unique in it's ability to act in response to a specific DNA sequence?

As a computer scientist, I'm interested in the ability of the Cas9 protein to function as an if-gate for DNA. It opens up so many questions for me. Right now the protein works as "if the sequence ...
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What´s the role or function of the homologous arms in a donor template in a knockout/knock edition via Crispr-cas9?

I have to make an exposition in my university about Crispr-cas9 edition and I have some questions about the method. In the knock out/knock in technique is used a plasmid containing the DNA that ...
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How to edit/insert new gene after cutting with CRISPR/cas9

I'm a student started who has started learning about CRISPR/Cas9. As I understand it, CRISPR/Cas9 is an enzyme that is used to cut a gene at a specific sequence. I would like to know how scientist do ...
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Do users of CRISPR/Cas iterate or parallelize to try multiple guide sequences?

I've read about on-target efficiency and off-target effects in use of CRISPR/Cas9, and about tools that suggest good guide sequences. I am wondering: how many guide sequences do typical CRISPR users ...
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How specific are CRISPR-cas9 cuts?

CRISPR-cas9 uses a string of RNA that matches with DNA and makes a double stranded cut at that point. If the RNA is just a few letters in length, the enzyme would cut DNA in many places. It would be ...
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why selecting cells after gene editing with CRISPR Cas9?

I am new to the CRISPR Cas9 genome editing system and I have the above basic question. Another way to think about this: what would happen if after cells are transfected, the cultures are left to grow ...