AGBE: a dual deaminase-mediated base editor by fusing CGBE with ABE for creating a saturated mutant population with multiple editing patterns

Yanhui Liang,Jingke Xie,Quanjun Zhang, Xiaomin Wang1,Shixue Gou1,Lihui Lin1,Tao Chen,Weikai Ge1,Zhenpeng Zhuang,Meng Lian,Fangbing Chen1, Nan Li,ZhenOuyang,Chengdan Lai,Xiaoyi Liu1,Lei Li1,Yinghua Ye,HanWu,Kepin Wang,Liangxue Lai

Nucleic Acids Research

2022, 50, 9, 5384-5399

Liangxue Lai

19.160

 Establishing saturated mutagenesis in a specific gene through gene editing is an efficient approach for identifying the relationships between mutations and the corresponding phenotypes. CRISPR/Cas9-based sgRNA library screening often creates indel mutations with multiple nucleotides. Single base editors and dual deaminase-mediated base editors can achieve only one and two types of base substitutions, respectively. A new glycosylase base editor (CGBE) system, in which the uracil glycosylase inhibitor (UGI) is replaced with uracil-DNA glycosylase (UNG), was recently reported to efficiently induce multiple base conversions, including C-to-G, C-to-T and C-to-A. In this study, we fused a CGBE with ABE to develop a new type of dual deaminase-mediated base editing system, the AGBE system, that can simultaneously introduce 4 types of base conversions (C-to-G, C-to-T, C-to-A and A-to-G) as well as indels with a single sgRNA in mammalian cells. AGBEs can be used to establish saturated mutant populations for verification of the functions and consequences of multiple gene mutation patterns, including single-nucleotide variants (SNVs) and indels, through high-throughput screening.