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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 ...

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Computing a function of DNA via crisppr-cas9 [on hold]

I wonder, whether the capabilities of Crisper-cas9 is to transform a given chain of DNA ( let call it x, where $x=[A,G,C,T]^*$). is it possible to encode the crisper as a function of x, and then the ...
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1answer
18 views

What is the role of CRISPR-dCas9 in gRNA-dCas9 transcription regulator complexes?

In this paper, 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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1answer
28 views

Are Restriction Enzymes obsolete with CRISPR?

Are Restriction Enzymes obsolete with CRISPR?
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39 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 ...
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1answer
31 views

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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2answers
26 views

How was Crisper Cas9 function discovered?

What paper outlines the discovery of how the CRIPER DNA, CrRNA, and Cas9 interact?
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16 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 ...
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1answer
4k 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: ...
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31 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 ...
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39 views

CRISPR/Cas9 in E Coli: whole gene insertion

This is a long shot, but can someone explain/refer me to a good explanation about how to design plasmids for whole gene insertion using CRISPR/Cas9 in E. coli? I've looked all over Addgene and read a ...
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29 views

Any idea about CRISPR-based gene activation in plants using p300 Acetyltransferase or Tet1 Demethylase?

We are planning to use CRISPR tools as RNP for epigenetic studies in plants. To this end, I'm wondering if bacterial production of dCas9 fused to the catalytic domain of p300 acetyltransferase or Tet1 ...
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1answer
60 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 ...
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50 views

Role of homology arms in CRISPR

I'm not a biologist so I am really confused about crispr... I am confused as to whether or not the homology arms of the donor oligonucleotide are what is placed inside the target DNA strand to replace ...
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27 views

How to design a primer for my T7 assay to validate my edit using CRISPR?

I have electroporated my cells with my CRISPR/Cas9 complex and now want to validate my edit using the T7 assay. Is there some tool I can use to find the best primers for the CRISPR sequence?
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2answers
60 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 ...
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0answers
36 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 ...
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1answer
145 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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30 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 ...
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16 views

Raising the monosodium glutamate content in tomatoes via Crispr

Is it possible to boost the monosodium glutamate content of tomatoes via Crispr? If so, can someone point me to some good reading materials/referenes on this subject? Thanks,
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94 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 ...
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1answer
133 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.
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1answer
44 views

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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2answers
40 views

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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1answer
34 views

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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3answers
5k views

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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1answer
222 views

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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60 views

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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1answer
111 views

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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33 views

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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1answer
187 views

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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1answer
136 views

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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2answers
722 views

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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2answers
6k views

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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2answers
46 views

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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506 views

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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2answers
4k views

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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0answers
63 views

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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1answer
115 views

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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169 views

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 ...
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1answer
171 views

What is the most up-to-date CRISPR/Cas9 protocol?

I am collecting literature to start a new project on CRISPR/Cas9 gene editing. I must put together a protocol to start and am intending to use the following paper as guidance: "Genome engineering ...
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74 views

Insertion of synthetic DNA sequence

If I have some synthetic DNA sequence (<=20 bp long), is there a way for me to reliably insert this sequence next to some n-bp motif? I'd like for this to be possible in humans. If so, are there ...
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1answer
631 views

In CRISPR bacteria, how does viral genomes get integrated into the spacers of CRISPR? Also, in its use, where does Cas9 cut the DNA?

I've been out of Biology for about a year polishing my programming skills. I know CRISPR/Cas9 allows targeted 'cutting' of DNA via RNA-guidance. Few questions regarding this. Regarding to its natural ...
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1answer
781 views

How does one insert cas9 into animal cells?

How could cas9 be inserted into cells by researchers looking to edit a genome? I imagine for engineering bacterial systems you could just put in the cas9 coding region in an expression vector, but is ...
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69 views

Why is dsDNA nuclease activity by CRISPR/Cas9 only shown indirectly?

In Jinek et al., the authors show nuclease activity of their CRISPR/Cas9 system using the so-called Surveyor assay method. This assay recognizes small mismatches in dsDNA which are introduced by error ...
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168 views

CRISPR-Cas Systems

In the context of the bacterial systems (not the gene editing tool), I was wondering what happens to the foreign DNA after the Cas proteins have created a new spacer. It is really not clear to me, ...
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1answer
215 views

Mutations/deletions with CRISPR

I need to stop some protein from being active and searching for some universal way to do so. In mammalians. With CRISPR it is possible to knock-out the entire gene. But it's a little complicate (...
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3answers
437 views

Splice in with CRISPR/Cas

I need to splice a gene into a human cell genome, with highest rate possible. I mean, doesn't really matter where the gene enters, nor does it matter if some cells die as a result of this. CRISPR ...
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0answers
91 views

CRISPR Knock in

Using the CRISPR/Cas9 technology, it is possible that after inducing a DSB with the Cas9 endonuclease guided with an RNA designed by the user and using a template DNA, get a desired Knock-In (KI) by ...