Questions tagged [cas9]
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CRISPR-Cas9 system, DNA repair
As a critical stage in the CRISPR-Cas9 system, two different mechanisms of DNA repair can occur in the target DNA after RNA has been introduced: non-homologous end joining (NHEJ) and homologous ...
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CRISPR/Cas9: How can inserted DNA be used as a donor for the homology-directed repair if Cas9 only creates blunt ends?
The CRISPR/Cas9 method allows new genes to be inserted. After Cas9 cuts the Target-DNA, it can use a homologous piece of DNA as a donor template for homology-directed repair. But HDR only occurs when ...
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Can the genetic sequences for CRISPR components be inserted into the host genome so that the cell perpetually produces CRISPR components?
Theoretically speaking, can you insert the gene sequences for cas9, sgRNA, and promoters into the host genome so that the cell perpetually produces CRISPR components?
In this scenario, I'm guessing ...
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How are spacer sequences created in a prokaryotic genome?
The CRISPR/Cas defense mechanism uses spacer sequences between palindromic repeats to search for the sequence to cut by an endonuclease. But how are these spacers created?
Let's take Bacteriophages, ...
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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 ...
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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 ...
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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 ...
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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 ...
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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 ...
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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-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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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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Can a protein go outside of nucleus after go inside with nuclear localization signal (NLS)
To make a protein (for example Cas9, GFP...) able to enter the nucleus, we need to add a NLS tag for it. So will it able to go outside the nucleus after get inside ?
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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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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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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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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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