DOI: 10.1002/ame2.70107
Generating golden Syrian hamsters with conditional alleles via zygote microinjection of CRISPR/Cas9
Wei Chen1
Xu Zhang1
Rui Fan2
Xia Li3
Feifei Guan1
Gefan Wan1
Weining Kong1
Xiaolong Qi1
Shuo Pan1
Sijing Shi2
Yuanlong Su2
Shan Gao4
Wei Huang2
Xunde Xian2
Jiangning Liu1
Yuhui Wang2
Yuanwu Ma5
1.State Key Laboratory of Respiratory Health and Multimorbidity,NHC Key Laboratory of Human Disease Comparative Medicine,Key Laboratory of Pathogen Infection Prevention and Control,Ministry of Education,National Human Diseases Animal Model Resource Center and National Center of Technology Innovation for Animal Model,Institute of Laboratory Animal Science,Chinese Academy of Medical Sciences(CAMS),Peking Union Medical College(PUMC),Beijing,China2.Institute of Cardiovascular Sciences,State Key Laboratory of Vascular Homeostasis and Remodeling,School of Basic Medical Sciences,Peking University,Beijing,China3.Department of Laboratory Animal Science,Peking University Health Science Center,Beijing,China4.State Key Laboratory of Respiratory Health and Multimorbidity,NHC Key Laboratory of Human Disease Comparative Medicine,Key Laboratory of Pathogen Infection Prevention and Control,Ministry of Education,National Human Diseases Animal Model Resource Center and National Center of Technology Innovation for Animal Model,Institute of Laboratory Animal Science,Chinese Academy of Medical Sciences(CAMS),Peking Union Medical College(PUMC),Beijing,China;Haihe Laboratory of Cell Ecosystem,Tianjin,China5.State Key Laboratory of Respiratory Health and Multimorbidity,NHC Key Laboratory of Human Disease Comparative Medicine,Key Laboratory of Pathogen Infection Prevention and Control,Ministry of Education,National Human Diseases Animal Model Resource Center and National Center of Technology Innovation for Animal Model,Institute of Laboratory Animal Science,Chinese Academy of Medical Sciences(CAMS),Peking Union Medical College(PUMC),Beijing,China;Haihe Laboratory of Cell Ecosystem,Tianjin,China;Medical Primate Research Center,Chinese Academy of Medical Sciences,Beijing,China
摘要：Background:The golden Syrian hamster is a valuable animal model for studying car-cinogenesis,metabolic disorders,cardiovascular diseases,and viral infections due to its biological and pathological similarities to humans.However,the development of ge-netically engineered hamsters has lagged behind that of mice and rats,largely because of an embryonic development block at the two-cell stage in vitro.Although CRISPR/Cas9-mediated gene knockout has been achieved in hamsters,precise DNA fragment insertion or conditional knockout(cKO)models have not previously been reported,likely due to technical limitations in embryo manipulation and insufficient efficiency of homology-directed repair(HDR).
Methods:In this study,we generated conditional alleles of the ApoF gene in golden Syrian hamsters.A two-cut strategy was applied using Cas9 protein,two sgRNAs,and a single donor plasmid containing exon 2 flanked by loxP sites and two～0.8 kb homol-ogy arms.A mixture of Cas9 protein,sgRNAs,and the donor plasmid was microin-jected into the pronuclei of one-cell stage hamster embryos.
Results:The efficiency of CRISPR/Cas9-mediated loxP knock-in reached up to 27％,and the genetically modified floxed alleles were successfully transmitted through the germline.The functionality of the inserted loxP sites was validated by in vivo Cre-mediated recombination following local administration of AAV vectors,including AAV-cTnT-Cre in the heart and AAV-CMV-Cre in the brain.
Conclusions:To our knowledge,this work represents the first successful establish-ment of a conditional knockout model in the golden Syrian hamster,providing a valu-able tool for mechanistic studies of gene function and disease modeling.
机标关键词：goldencrisprwithallelesconditionalgeneratinghamstersmicroinjection
论文发表日期：2026-02-28
在线出版日期：2026-03-31（本平台首次上网日期，不代表文献的发表时间）
页数：11( 308-318 )
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