CATI:通过MMEJ抑制对啮齿动物和灵长类动物胚胎进行有效的基因整合方法的数据表

同源定向修复(homology-directed repair, HDR)的效率在动物模型和基因治疗的发展中起着至关重要的作用。我们证明了微同源介导的末端连接(MMEJ)在crispr介导的基因编辑过程中构成了相当大的DNA修复比例。利用CasRx下调一个关键的MMEJ因子聚合酶Q (Polq)，我们提高了小鼠胚胎和后代中线性化DNA片段和单链寡核苷酸(ssODN)的靶向整合效率。casrx辅助靶向整合(CATI)也导致CRISPR/Cas9编辑猴胚胎过程中HDR效率的显著提高。我们提出了一种有前途的工具，用于生成猴子模型和开发临床试验的基因疗法。

The efficiency of homology-directed repair (HDR) plays a critical role in the development of animal models and gene therapy. We demonstrate that microhomology-mediated end joining (MMEJ) accounts for a substantial proportion of DNA repair events during CRISPR-mediated gene editing. By downregulating polymerase Q (Polq), a key MMEJ factor, via CasRx, we enhanced the targeted integration efficiency of linearized DNA fragments and single-stranded oligodeoxynucleotides (ssODN) in mouse embryos and their progeny. CasRx-assisted targeted integration (CATI) also resulted in a significant improvement in HDR efficiency during CRISPR/Cas9 editing of monkey embryos. We present a promising tool for generating monkey models and developing gene therapies for clinical trials.