The CRISPR/Cas9 enabled efficient gene editing in an easy and programmable manner. Controlling its activity in greater precision is desired for biomedical research and potential therapeutic translation. Here, we engrafted the CRISPR/Cas9 system with a mutated human estrogen receptor (ER^T2), which renders it 4-hydroxytamoxifen (4-OHT) inducible for the access of genome, and a nuclear export signal (NES), which lowers the background activity. Tight and efficient drug-inducible genome editing was achieved across several human cell types, including embryonic stem cells (ESCs) and mesenchymal stem cells (MSCs), upon vigorous optimization. Optimized terminal device, which we named hybrid drug inducible CRISPR/Cas9 technology (HIT-Cas9), delivered advantageous performances over several existing designs. Such architecture was also successfully applied to an orthogonal Cas9. The HIT-Cas9 system developed in this study will find broad utility in controlled editing of potentially any genomic loci.

CRISPR / Cas9以简单且可编程的方式实现了有效的基因编辑。生物医学研究和潜在的治疗翻译需要更精确地控制其活性。在这里，我们将CRISPR / Cas9系统植入了突变的人类雌激素受体（ER ^T2），从而使其可诱导4-羟基他莫昔芬（4-OHT）进入基因组，并产生核输出信号（NES），从而降低了背景活性。通过大力优化，可以在包括胚胎干细胞（ESC）和间充质干细胞（MSC）在内的几种人类细胞类型中实现紧密有效的药物诱导基因组编辑。经过优化的终端设备（我们称为混合药物诱导性CRISPR / Cas9技术（HIT-Cas9））在几种现有设计上均具有优越的性能。这种架构也成功地应用于正交Cas9。在这项研究中开发的HIT-Cas9系统将在可能编辑任何基因组基因座的受控编辑中找到广泛的用途。