A-to-G/C/T and C-to-T/G/A dual-function base editor for creating multi-nucleotide variants  

作　　者：Bingxiu Ma Han Wu Shixue Gou Meng Lian Cong Xia Kaiming Yang Long Jin Junyuan Liu Yunlin Wu Yahai Shu Haizhao Yan Zhanjun Li Liangxue Lai Yong Fan 

机构地区：[1]Department of Obstetrics and Gynecology,Guangdong Provincial Key Laboratory of Major Obstetric Diseases,Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology,Guangdong-Hong Kong-Macao Greater Bay Area Higher Education Joint Laboratory of Maternal-Fetal Medicine,The Third Affiliated Hospital of Guangzhou Medical University,Guangzhou,Guangdong 510150,China [2]China-New Zealand Joint Laboratory on Biomedicine and Health,CAS Key Laboratory of Regenerative Biology,Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine,Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou,Guangdong 510530,China [3]Sanya Institute of Swine Resource,Hainan Provincial Research Centre of Laboratory Animals,Sanya,Hainan 572000,China [4]Research Unit of Generation of Large Animal Disease Models,Chinese Academy of Medical Sciences(2019RU015),Guangzhou,Guangdong 510530,China [5]Guangzhou National Laboratory,Guangzhou,Guangdong 510005,China [6]Institute of Laboratory Animal Sciences,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100020,China [7]Key Laboratory of Zoonosis Research,Ministry of Education,College of Animal Science,Jilin University,Changchun,Jilin 130062,China

出　　处：《Journal of Genetics and Genomics》2024年第12期1494-1504,共11页遗传学报（英文版）

基　　金：supported by the National Key Research and Development Program of China(2021YFC2700904,2021YFF0702601,2021YFA0805300);the National Natural Science Foundation of China(82071723,82100482,82101733);Guangdong Basic and Applied Basic Research Foundation(2021B1515020069);Guangzhou Science and Technology Planning Project(2023A03J0377);Research Unit of Generation of Large Animal Disease Models,Chinese Academy of Medical Sciences(2019-I2M-5-025)。

摘　　要：Multi-nucleotide variants(MNVs)are critical genetic variants associated with various genetic diseases.However,tools for precisely installing MNVs are limited.In this study,we present the development of a dual-base editor,BDBE,by integrating TadA-dual and engineered human N-methylpurine DNA glycosylase(eMPG)into nCas9(D10A).Our results demonstrate that BDBE effectively converts A-to-G/C/T(referred to as A-to-B)and C-to-T/G/A(referred to as C-to-D)simultaneously,yielding nine types of dinucleotides from adjacent CA nucleotides while maintaining minimal off-target effects.Notably,BDBE4 exhibits exceptional performance across multiple human cell lines and successfully simulated all nine dinucleotide MNVs from the gnomAD database.These findings indicate that BDBE significantly expands the product range of base editors and offers a valuable resource for advancing MNV research.

关 键 词：CRISPR Gene editing Dual-base editor A-to-G/C/T and C-to-T/G/A MNV Genetic diseases 

分 类 号：Q78[生物学—分子生物学]