I wish to know this for the feasibility and reliability as well as accuracy.
this is arguably an unanswerably question, as a deep understanding of nuclease-mediated gene editing will expose slight advantages to any system. however, even without knowing your model or question, I suspect -with reasonable confidence- that you will end up employing a Cas9 approach. i say this due to the relative ease of use, cost of use and reliability of results. for example if your cell is amenable to transfection by electroporation or lipofect then you could go from design to bialleliac knockout within several weeks with a cost largely dependent on culture media and sorting reagents. the later of these is not to be overlooked: developing a reliable sorting strategy to identify mutants will obviate the need for high efficiency at the genome level. i.e. if you have a selectable marker such as an antibody for flow-sort you can get by with very low mutation rates. alternatively you may rely on genomic screening assays such as the t7 or simply sanger sequencing.
hope this helps.
all the best,
James
Article Recent Advances in Genome Editing and Creation of Geneticall...
Thanks for the article. Its awesome and very much informative. I got through the comparative analysis and i am impressed.
I would suggest Double Nickase strategy with the CRISPR-cas9 (D10 mutant). easy to clone gRNAs to any target region and quite specific since you need 2 separate Cas9 molecules to bind to your target area and realize DNA cleavage. the regular Cas9 is therefore able to generate more off-taget double strand brakes. just have a look at all the protocols at www.addgene.com
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