Generation of C-to-G transversion in mouse embryos via CG editors
Generation of C-to-G transversion in mouse embryos via CG editors
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DOI：
10.1007/s11248-022-00313-x
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发表时间：
2022-06-15
影响因子：
3
通讯作者：
Huang, Junjiu
中科院分区：
文献类型：
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作者：
Cao, Tianqi;Liu, Simiao;Huang, Junjiu
碱基编辑器（BE）是用于在活生物体中产生单碱基转换的高效且精确的工具。虽然大多数BE系统在介导C到T或A到G转换方面受到限制，但最近开发的C到G碱基编辑器（CGBE）可以产生C到G颠换。CGBE在编辑窗口内将胞嘧啶转化为无碱基中间体，其将在碱基切除修复（BER）后被任何碱基替换。到目前为止，尽管已经通过gRNA文库和机器学习在培养细胞中研究了CGBE的效率和编辑范围，但CGBE在生成小鼠模型中的可行性尚未得到充分测试。在这项研究中，我们测试了CGBE 1和CGBE-XRCC 1系统在小鼠胚胎中的C到G颠换效率。我们的结果表明，两种CGBE系统都能够在3个测试靶标中的2个上介导C至G颠换，在编辑窗口内频率高达20%。值得注意的是，大多数组显示出超过40%的其他碱基转化，主要是C至T。最后，我们成功获得了携带致病突变的F-1小鼠。总之，我们的研究表明，CGBEs系统在建立小鼠模型方面具有巨大的潜力，并表明基于XRCC 1的系统适用于小鼠胚胎。
Base editors (BEs) are efficient and precise tools for generating single base conversions in living organisms. While most BE systems are limited in mediating C-to-T or A-to-G conversions, recently developed C-to-G base editors (CGBEs) could produce C-to-G transversions. CGBEs convert cytosine within the editing window to abasic intermediates, which would be replaced with any base after base excision repair (BER). By far, though the efficiency and editing scope of CGBEs have been investigated in cultured cells via gRNA library and machine-learning, the viability of CGBEs in generating mouse models has not been adequately tested. In this study, we tested the C-to-G transversion efficiency of the CGBE1 and CGBE-XRCC1 systems in mouse embryos. Our results showed that both of the CGBE systems were able to mediate C-to-G transversion on 2 out of 3 targets tested, with up to 20% frequency within the editing window. Notably, most of the groups showed over 40% of other base conversions, predominantly C-to-T. Lastly, we successfully acquired the F-1 mouse carrying a disease-causing mutation. In all, our study suggested that CGBEs systems held great potential in generating mouse models and indicated that XRCC1 based system is applicable in mouse embryos.