Thursday, October 16, 2014

A review about #CRISPR off-target effects with links to 10 online off-target identification tools.

Gene editing: how to stay on-target with CRISPR.  Vivien Marx.  Nat Methods. 2014 Sep 29;11(10):1021-6. doi: 10.1038/nmeth.3108.

In addition, let me reiterate some rules of thumb about minimizing off-target effects:

1.  3 or more mismatches in the 20-base protospacer are highly likely to prevent off-target cleavage.

2.  Or, consider using truncated protospacers of 17 or 18 bases.  The catch is you will probably have to pick a protospacer with a G as the first base to allow guide RNA transcriptional initiation.

3. I'm not sure if paired nickases are the best way to go yet.  A better way to think of it is that it may depend on the specific goal of your CRISPR experiment.      

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