Wei Yu, Zhang Xiao Hui, Li Da Li
Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai 200241, China.
Yi Chuan. 2017 Dec 20;39(12):1115-1121. doi: 10.16288/j.yczz.17-389.
Programmable nucleases are cutting edge genetic technology which edits targeted DNA sequences through generation of site-specific double-strand DNA breaks (DSBs). To improve the efficiency and precision of genetic modification, scientists have developed a single-base editing system (base editor) through combining of CRISPR/Cas9 system with cytosine deaminase. Compared with Cas9 system, this base editor can convert cytosine to thymine (C > T) at specific site more efficiently without inducing DSBs to avoid generation of indels. However, the base editor can only generate transition of pyrimidine but could not modify purines. Recently, Nature published a novel base editing system to convert adenine to guanine (ABEs, adenine base editors) through fusion of Cas9 nickase to a modified deaminase which is evolved through screening of random library based on tRNA adenine deaminase from E. coli. Here, we summarize the development of single-base editing tools and the latest research progress, especially the optimization process of ABEs, as well as the potential directions of the base editors.
可编程核酸酶是前沿的基因技术,它通过产生位点特异性双链DNA断裂(DSB)来编辑靶向DNA序列。为了提高基因编辑的效率和精度,科学家们通过将CRISPR/Cas9系统与胞嘧啶脱氨酶相结合,开发出了一种单碱基编辑系统(碱基编辑器)。与Cas9系统相比,这种碱基编辑器能够在特定位点更高效地将胞嘧啶转化为胸腺嘧啶(C>T),而不会诱导DSB产生,从而避免了插入缺失的产生。然而,该碱基编辑器只能产生嘧啶转换,无法修饰嘌呤。最近,《自然》杂志发表了一种新型碱基编辑系统,通过将Cas9切口酶与一种经过修饰的脱氨酶融合,该脱氨酶是基于大肠杆菌的tRNA腺嘌呤脱氨酶通过随机文库筛选进化而来,从而将腺嘌呤转化为鸟嘌呤(ABEs,腺嘌呤碱基编辑器)。在此,我们总结了单碱基编辑工具的发展和最新研究进展,特别是ABEs的优化过程,以及碱基编辑器的潜在发展方向。