Cas9_cutting_efficiency

The CRISPR system is becoming more popular as a tool for genome modification in mammalian cells, expecially in human induced pluripotent stem (iPS) cells and embryonic stem (ES) cells. The efficiencies of double-strand break (DSB) introduction by the CRISPR system are key for the successful engineering of the iPS/ES cell genome. To use custom Cas9 cassettes, we need to measure off-target "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" efficiency measurement. We expect that one run of MiSeq should be albe to analyze efficiency of more than 100 Cas9 cassettes.