| Autor: |
Yanhui Liang, Jingke Xie, Quanjun Zhang, Xiaomin Wang, Shixue Gou, Lihui Lin, Tao Chen, Weikai Ge, Zhenpeng Zhuang, Meng Lian, Fangbing Chen, Nan Li, Zhen Ouyang, Chengdan Lai, Xiaoyi Liu, Lei Li, Yinghua Ye, Han Wu, Kepin Wang, Liangxue Lai |
| Rok vydání: |
2022 |
| Předmět: |
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| Zdroj: |
Nucleic acids research. 50(9) |
| ISSN: |
1362-4962 |
| Popis: |
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. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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