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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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-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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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 ...
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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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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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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 ...
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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 ...
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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 ...
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78 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?
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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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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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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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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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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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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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Do single oligo sgRNAs for CRISPR require a modified CAS9?

I am attempting to use CRISPR to insert a tag into a gene. The way we have done this in the past is to use separate crRNA (with a sequence to interact with the tracr) and the universal Tracr sequence ...
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Can base editors be anything else than SpCas9 and SaCas9 variants?

I found out that only variants of Streptococcus pyogenes Cas9 (SpCas9) and Staphylococcus aureus Cas9 (SaCas9) are used with adenine or cytidine base editors, and I was wondering why. Can Neisseria ...
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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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202 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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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 ...