Exome sequencing to detect on- and off-target editing using self-inactivating CRISPR (SiC) system

A SiC vector system was developed. A pool of 5 lentivirus containing sgRNA targeting specific genes was developed and transduced into HEK-Cas9dTomato-Cas9G7 cells. In some cases, Doxycycline (Dox) was added at various times (-18h before transduction, 0 day at the time of transduction, +2 days after transduction). In one case Dox was absent [positive control], and in another control the cells were untransduced [negative control]. Exome sequencing was performed using genomic DNA collected on day 15.