Efficient base editing for multiple genes and loci in pigs using base editors
Efficient base editing for multiple genes and loci in pigs using base editors
复制标题
使用碱基编辑器对猪的多个基因和基因座进行高效碱基编辑
DOI：
10.1038/s41467-019-10421-8
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发表时间：
2019-06-28
影响因子：
16.6
通讯作者：
Lai, Liangxue
中科院分区：
文献类型：
--
作者：
Xie, Jingke;Ge, Weikai;Lai, Liangxue
胞嘧啶碱基编辑器（CBE）能够在多种生物体中实现可编程的C-T转换而无需DNA双链断裂和同源定向修复，这在农业和生物医学应用中表现出巨大的潜力。然而，所有报告的病例仅涉及单个靶向基因组位点的C至T取代。在大型动物中，尚未验证C至T取代是否在多个位点/基因座中有效。在这里，通过使用猪，一种重要的农业和生物医学动物，作为主体动物，我们表明CBE可以有效地诱导多个位点/位点的C-到-T转换与三个基因的组合，包括DMD，TYR，和LMNA，或RAG 1，RAG 2，和IL 2 RG，同时，在胚胎和细胞水平。CBE还可以通过引入多个提前终止密码子破坏数十个拷贝的基因（猪内源性逆转录病毒pol基因）。利用CBE，通过胚胎注射或核移植方法产生携带单基因或多基因点突变的猪。
Cytosine base editors (CBEs) enable programmable C-to-T conversion without DNA double-stranded breaks and homology-directed repair in a variety of organisms, which exhibit great potential for agricultural and biomedical applications. However, all reported cases only involved C-to-T substitution at a single targeted genomic site. Whether C-to-T substitution is effective in multiple sites/loci has not been verified in large animals. Here, by using pigs, an important animal for agriculture and biomedicine, as the subjective animal, we showed that CBEs could efficiently induce C-to-T conversions at multiple sites/loci with the combination of three genes, including DMD, TYR, and LMNA, or RAG1, RAG2, and IL2RG, simultaneously, at the embryonic and cellular levels. CBEs also could disrupt genes (pol gene of porcine endogenous retrovirus) with dozens of copies by introducing multiple premature stop codons. With the CBEs, pigs carrying single gene or multiple gene point mutations were generated through embryo injection or nuclear transfer approach.