gRNA_Cas9_validation-sc-2013-07-25T12:05:16Z-2711
gRNA_Cas9_validation
PCR products are obtained from each target loci using genomic DNA from gRNA/Cas9-treated 293T cells. Subsequently, PCR products are pooled and subjected to Illumina library preparation and Miseq sequencing. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
gRNA/Cas9 system is becoming more general tools for genome modification in mammalian cells, such as in human 293T cells, K562 cells, induced pluripotent stem (iPS) cells and embryonic stem (ES) cells. The efficiencies of double-strand break (DSB) introduction by gRNA/Cas9 are key for the successful engineering of the cell genome. To use gRNA/Cas9 system, we need to measure "cutting" efficiency by a precise method, which can be adapted in a high-throughput manner. In this study, we explore the possibility of MiSeq sequencing for the "cutting" efficinecy measurement. We expect that one run of MiSeq should be able to analyze efficiency of more than 12 gRNA/Cas9