Indel profiling of primary hepatocytes by deep sequencing

CRISPR/Cas9 has proven to be an efficient tool for gene editing both in vitro and in vivo. The aim of this study is to evaluate editing efficiency of various genes targted in Rosa26-Cas9 knock-in mice. Efficiency of in vivo genome editing was assessed using for hepatocytes isolated from the livers of Cas9-EGFP mice to which adeno-assocaited viruses (AAVs) packaged with sgRNAs targeting various genes were injected. Genemic regions containing each cut site were amplified by PCR, and libraries were prepared for deep sequencing.

CRISPR/Cas9已被证实为一种可用于体外（in vitro）与体内（in vivo）基因编辑的高效工具。本研究旨在评估靶向于Rosa26-Cas9敲入（knock-in）小鼠的各类基因的编辑效率。本研究以从Cas9-EGFP小鼠肝脏中分离的肝细胞为样本评估体内基因组编辑效率，此类小鼠已被注射了包装有靶向各类基因的单导RNA（single guide RNAs, sgRNAs）的腺相关病毒（adeno-associated viruses, AAVs）。将包含每个切割位点的基因组区域通过聚合酶链式反应（polymerase chain reaction, PCR）扩增，并构建文库用于深度测序。