Is it possible that the mutation did not occur on the guided RNA spacer sequence instead in different area closer to the gRNA spacer sequence?

23 April 2015 1 4K Report

I know that the cut or the DSB happened 5-6bp away from the PAM sequence, inside the guided RNA spacer sequence. But is it possible that the mutation occurred not in the guided RNA spacer sequence? Thanks.

Subin R. C. K. Rajendran

I don't fully understand the quetion you are asking. Are asking whether Cas9 can cut at other locations beside the gRNA target sequence? Spacers are associated with the native CRISPR loci in bacteria where the repeats are interspersed by the spacers. For a non native application of CRISPR system in a different species. I would think it is more appropriate to call it as gRNA target sequence. If you could elaborate or clarify, that would be good.

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