When working with CRISPR approaches all kind of mutations can occur through NHEJ. Most frequently I and others see small (1-5nt) deletions, less frequently insertions, sometimes lager deletions. The biggest deltion I have found recently on a mouse ES cell projest is 350 nt. In a recent report from Alan Bradley () a much bi...
I am trying to screen for gRNA's targeting a plasmid that is being co-transfected into HEK 293T cells. However, when I sequence around the cut site for TIDE, I am not seeing any detectable indel formations. Has anyone done this before? Can plasmids undergo NHEJ, or is it that the plasmids undergo degradation after cutting?...
I did a literature search tailored to find ways to increase HDR in Human IPS - ES cells. I was interested in trying drugs to enhance Homology directed repair. Has anyone has tried any of the following drugs- constructs in their lab or know of any which I have left off the list. I will be introducing a SNP into an IPSC ...
Using the protocol outlined below, I am unable to detect integration of our donor construct by PCR or GFP expression in THP-1 cells. Does anyone have a protocol or a reference to the use of recombinant Cas9 protein in THP-1 cells using nucleofection, or a suggestion to improve our current protocol? THP-1 cells are recalcitr...
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