Human 8-cell embryos enable efficient induction of disease-preventive mutations without off-target effect by cytosine base editor

作     者：Yinghui Wei Meiling Zhang Jing Hu Yingsi Zhou Mingxing Xue Jianhang Yin Yuanhua Liu Hu Feng Ling Zhou Zhifang Li Dongshuang Wang Zhiguo Zhang Yin Zhou Hongbin Liu Ning Yao Erwei Zuo Jiazhi Hu Yanzhi Du Wen Li Chunlong Xu Hui Yang Yinghui Wei;Meiling Zhang;Jing Hu;Yingsi Zhou;Mingxing Xue;Jianhang Yin;Yuanhua Liu;Hu Feng;Ling Zhou;Zhifang Li;Dongshuang Wang;Zhiguo Zhang;Yin Zhou;Hongbin Liu;Ning Yao;Erwei Zuo;Jiazhi Hu;Yanzhi Du;Wen Li;Chunlong Xua;Hui Yang

作者机构：Center for Reproductive MedicineInternational Peace Maternity and Child Health HospitalInnovative Research Team of High-level Local Universities in ShanghaiSchool of MedicineShanghai Jiao Tong UniversityShanghai 2000301.Center for Reproductive MedicineInternational Peace Maternity and Child Health HospitalInnovative Research Team of High-level Local Universities in ShanghaiSchool of MedicineShanghai Jiao Tong UniversityShanghai 200030China Institute of NeuroscienceState Key Laboratory of NeuroscienceKey Laboratory of Primate NeurobiologyCAS Center for Excellence in Brain Science and Intelligence TechnologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China Lingang LaboratoryShanghai Research Center for Brain Science and Brain-Inspired Intelligence TechnologyShanghai 200031China Center for Reproductive MedicineRen Ji HospitalSchool of MedicineShanghai Key Laboratory for Assisted Reproduction and Reproductive GeneticsShanghai Jiao Tong UniversityShanghai 200127China The MOE Key Laboratory of Cell Proliferation and DifferentiationSchool of Life SciencesGenome Editing Research CenterPeking UniversityBeijing 100871China Center for Reproductive MedicineShandong UniversityJinan 250012China Center for Reproductive MedicineAnhui Provincial Maternal and Child Health HospitalHefei 230001China Shenzhen BranchGuangdong Laboratory for Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518000China Reproductive Medicine CenterDepartment of Obstetrics and Gynecologythe First Affiliated Hospital of Anhui Medical UniversityHefei 230022China 

出 版 物：《Protein & Cell》 (蛋白质与细胞（英文版）)

年 卷 期：2023年第14卷第6期

页      面：416-432页

核心收录：

学科分类：10[医学] 

基　　金：supported by Chinese National Science and Technology major project R&D Program of China(2018YFC2000101) Strategic Priority Research Program of Chinese Academy of Science(XDB32060000) National Natural Science Foundation of China(Grant Nos.31871502,31901047,31925016,91957122,82021001,and 31922048) Basic Frontier Scientific Research Program of Chinese Academy of Sciences From 0 to 1 original innovation project(ZDBS-LYSM001) Shanghai Municipal Science and Technology Major Project(2018SHZDZX05) Shanghai City Committee of Science and Technology Project(18411953700,18JC1410100,19XD1424400 and 19YF1455100) Innovative Research Team of High-Level Local Universities in Shanghai(SHSMU-ZDCX20212200 and SHSMU-ZLCX20210200) International Partnership Program of Chinese Academy of Sciences(153D31KYSB20170059) Postdoctoral Science Foundation of China(2020M681417 and 2021T140684) Sailing Program of Shanghai(21YF1453000)(to J.H.) 

主　　题：human embryo APOE4 disease-preventive mutations base editor 

摘      要：Approximately 140 million people worldwide are homozygous carriers of APOE4(ε4),a strong genetic risk factor for late onset familial and sporadic Alzheimer’s disease(AD),91%of whom will develop AD at earlier age than heterozygous carriers and *** to AD could be reduced by targeted editing of APOE4,but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene ***,we first screened eight cytosine base editor variants at four injection stages(from 1-to 8-cell stage),and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate(up to 100%)with the lowest bystander *** particular,80%of AD-susceptibleε4 allele copies were converted to the AD-neutralε3 allele in humanε4-carrying *** control measures combined with targeted deep sequencing,whole genome sequencing,and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem ***,base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst ***,we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial *** study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos,a potential approach for reducing human susceptibility to AD or other genetic diseases.