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.

Filter by
Sorted by
Tagged with
3 votes
0 answers
45 views

Is CRISPR mediated RNA editing less specific and less efficient than DNA editing?

According to this diagram, the high efficiency and the high specificity of CRISPR lies in its reversible binding with the target DNA. The Cas protein unzips the target DNA and have the gRNA to base ...
user avatar
  • 393
0 votes
0 answers
30 views

What will be the effect of "gene-drives" on natural selection and speciation?

I understand that gene-drives not only contain a genetic mutation brought about by CRISPR-CAS, but that rather they also contain the alleles necessary for CRISPR-cas to operate in descendants. Thus ...
user avatar
  • 121
0 votes
0 answers
39 views

What is the fastest way to find all 23-mers in the human genome (GRCh38) that end with NGG?

According to this paper, there are 0.68 billion such sequences. The authors have mentioned that the data was collected from ENCODE. But I could not find any sequence download link on that site. So far,...
user avatar
1 vote
0 answers
34 views

Would viral diversity result in a change in the effectiveness of CRISPR systems in a population of bacteria, within a closed system?

I have here my hypothesis, does this make scientific sense? Assume this situation is occurring in a closed environment with only bacteria and bacteriophages. The effectiveness of CRISPR/Cas9, being an ...
user avatar
  • 19
0 votes
1 answer
32 views

Do bacteria duplicate the copied virus DNA to put into a Cas9 protein when fighting the virus off again?

When bacteria insert a part of invading virus DNA into its own genetic sequence, on the 2nd invasion does it duplicate that copied DNA again from wherever the bacteria placed it and put it into a Cas9 ...
user avatar
0 votes
1 answer
54 views

Can CRISPR knockout of a gene be referred gene inhibition?

Can gene knockout (e.g. by CRISPR) be referred to as gene inhibition? I am trying to use precise words for a manuscript. We first showed siRNA of a gene has effect X. We also found knockout of the ...
user avatar
  • 111
-2 votes
1 answer
88 views

Can we cure cancer with CRISPR dead Cas?

Here's a silly idea I had this morning: Sequence a bunch of normal patient cells. Sequence a bunch of tumor cells from a biopsy. Find a DNA sequence that we're reasonably certain exists in the cancer ...
user avatar
3 votes
1 answer
126 views

Does ribonuclease processing of pre-crRNAs happen co-transcriptionally?

I understand CRISPR-mediated bacterial immunity to occur in the following simplified steps: A CRISPR array is transcribed from promoters in the leader sequence to yield a precursor CRISPR RNA (pre-...
user avatar
  • 7,564
1 vote
1 answer
55 views

Is CRISPR being utilized when scientists use the CRISPR/Cas9 system to edit genes?

"CRISPR" and "Cas9" are different things. When a virus attacks a bacteria, the bacteria stores the viral code of the virus in CRISPR. And when the virus attacks again the Cas9 ...
user avatar
  • 493
0 votes
0 answers
56 views

How was gene knock out done in pre CRISPR era?

I am trying to understand how CRISPR has made the gene knockout or gene editing process simpler to make transgenic animals. Here is an old (pre CRISPR) flowchart from Manis, 2007 that shows how ...
user avatar
  • 3,636
2 votes
0 answers
22 views

What obstacle(s) would be most prevalent in an attempt to use virotherapy to inject Cas into cells to target a cancerous mutation?

Let's say a human cell mutates cancerously, we identify the mutated gene sequence and use CRISPR to mutate the patient's cells to include some variation of the cas genes and spacer sequences matching ...
user avatar
  • 121
6 votes
1 answer
187 views

When does the Cas9 nuclease stop?

If I understand correctly, the steps of gene editing with CRISPR-Cas9 are roughly as follows Cas9 nuclease and guide RNA form a complex. Cas9+guide RNA complex scans genomic DNA and recognizes the ...
user avatar
1 vote
0 answers
86 views

if an SNP were edited using CRISPR What are the chances that, absent artificial selection, wild type alleles would reemerge?

I am researching a fatty acid amide hydrolase (FAAH) SNP RS324420 and FAAH out microdeletion that together lead to reduced pain sensitivity and reduced anxiety (Moreira et al 2008). The causative ...
user avatar
-1 votes
1 answer
171 views

Can we change the Eye/Hair color by knocking out the OCA2, HERC2 and MC1R genes using CRISPR in an adult human?

This paper seems to describe the use of a plasmid delivered by a gene gun to depigment rat skin; https://www.nature.com/articles/3302264 Published: 27 May 2004 Seeing the gene therapy: application of ...
user avatar
1 vote
1 answer
21 views

Does immunity to CRISPR proteins limit their effectiveness?

The use of crisper-cas systems is currently applied to cells cultivated in vitro. As control of the ‘off target’ effects of Crispr improves and Crispr is used in vivo, why won’t the immune system ...
user avatar
0 votes
1 answer
107 views

Gene knockdown vs gene knockout vs knocksideways? [closed]

How are the techniques: Knock-sideways, knockout & knock-down different?
user avatar
0 votes
2 answers
135 views

Can CRISPR be used in editing primary cells? How is the transfection efficiency?

Sometimes people wanna use primary cells to do gene-editing because the cells normally have more interesting characteristics, but can we really do so?
user avatar
1 vote
1 answer
56 views

Does using CRISPR/Cas9 knockout need to add donor DNA in the process?

I am new to this technology and don't quite understand how it works. Hope someone can give some suggestions! Thank you.
user avatar
0 votes
1 answer
57 views

simple explanation of a crispr screen vs pooled crispr screen?

I'm familiar with how CRISPR works to make either knockdown or amplification effects for a single gene because you can make precision cuts. What exactly does a pooled crispr screen do & how does ...
user avatar
2 votes
1 answer
85 views

CRISPR-Cas9 knockout

With CRISPR-Cas9 I have conducted a targeted knockout of a DNA region encoding a certain protein (working with leukocytes). My question is, how long does it take until this protein is not detectable ...
user avatar
  • 21
2 votes
1 answer
89 views

What software do I need to read-write cas9? and .dna files?

I am still a complete beginner to crispr and I am still trying to learn what it is and how to actually use it. I now realise that you have to order the crispr components after you have actually ...
user avatar
0 votes
1 answer
68 views

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? ...
user avatar
  • 15
0 votes
1 answer
108 views

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 ...
user avatar
4 votes
2 answers
305 views

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/...
user avatar
  • 153
0 votes
1 answer
83 views

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 ...
user avatar
  • 389
2 votes
1 answer
60 views

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 ...
user avatar
  • 21
2 votes
1 answer
366 views

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, ...
user avatar
0 votes
1 answer
68 views

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 ...
user avatar
1 vote
1 answer
143 views

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 ...
user avatar
-1 votes
2 answers
190 views

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 ...
user avatar
  • 231
0 votes
1 answer
36 views

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 ...
user avatar
1 vote
1 answer
37 views

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 ...
user avatar
1 vote
0 answers
88 views

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?
user avatar
  • 915
1 vote
0 answers
109 views

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 ...
user avatar
  • 212
1 vote
1 answer
50 views

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?
user avatar
  • 15
1 vote
1 answer
76 views

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 ...
user avatar
  • 389
1 vote
1 answer
246 views

Are Restriction Enzymes obsolete with CRISPR?

Are Restriction Enzymes obsolete with CRISPR?
user avatar
  • 1,637
1 vote
1 answer
237 views

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 ...
user avatar
  • 373
0 votes
1 answer
67 views

Does gene editing have any technical cross-species limits?

How far could you go in cross-species gene editing? Is it possible for example to introduce plant genes into human DNA and vice versa? Question: How is gene editing limited by the "donor" and "...
user avatar
  • 403
0 votes
2 answers
174 views

How was Crisper Cas9 function discovered?

What paper outlines the discovery of how the CRIPER DNA, CrRNA, and Cas9 interact?
user avatar
  • 1,637
1 vote
0 answers
53 views

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 ...
user avatar
34 votes
1 answer
5k views

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: ...
user avatar
  • 443
1 vote
0 answers
51 views

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 ...
user avatar
  • 111
2 votes
1 answer
98 views

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 ...
user avatar
  • 191
3 votes
2 answers
154 views

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 ...
user avatar
  • 1,261
1 vote
0 answers
48 views

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 ...
user avatar
1 vote
1 answer
323 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.
user avatar
1 vote
0 answers
58 views

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 ...
user avatar
  • 151
1 vote
0 answers
120 views

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 ...
user avatar
0 votes
1 answer
388 views

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.
user avatar