AGBE: a dual deaminase-mediated base editor by fusing CGBE with ABE for creating a saturated mutant population with multiple editing patterns. Issue 9 (11th May 2022)

• Record Type: Journal Article
• Authors: Liang, Yanhui; Xie, Jingke; Zhang, Quanjun; Wang, Xiaomin; Gou, Shixue; Lin, Lihui; Chen, Tao; Ge, Weikai; Zhuang, Zhenpeng; Lian, Meng; Chen, Fangbing; Li, Nan; Ouyang, Zhen; Lai, Chengdan; Liu, Xiaoyi; Li, Lei; Ye, Yinghua; Wu, Han; Wang, Kepin; Lai, Liangxue
• Is Part Of: Nucleic acids research. Volume 50:Issue 9(2022)
• Page Start: 5384
• Page End: 5399
• Publication Date: 2022-05-11
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gkac353
• Languages: English
• ISSNs: 0305-1048

Abstract

Abstract: 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.