ZFN_TALEN_validation_in_human_iPS_cells

Zinc finger nucleases (ZFNs) and TAL-effector nucleases (TALENs) are becoming more general tools 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 these neculeases are key for the successful engineering of the iPS/ES cell genome. To use custom ZFNs/TALENs, 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 albe to analyze efficinecy of more than 100 necleases. As a pilot stuy, we will use 8 ZFNs or TALENs we have got so far.