Generation and phenotypic analysis of Fancm gene knockout mice
Received:November 16, 2018  
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DOI:10. 3969 / j.issn.1005-4847. 2019. 03. 009
KeyWord:Fanconi anemia; FANCM ; CRISPR/ Cas9; Fancm gene knockout; testis development defect; mice
                       
AuthorInstitution
杨桥 1. 浙江省医学科学院,杭州 ; 2. 浙江中医药大学,杭州
郭红刚 浙江省医学科学院,杭州
楼琦 浙江省医学科学院,杭州
柯贤福 浙江省医学科学院,杭州
周文伟 浙江省医学科学院,杭州
应华忠 浙江省医学科学院,杭州
俞冰 浙江中医药大学,杭州
张婷婷 浙江省医学科学院,杭州
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Abstract:
      Objective To generate a Fancm gene knockout mouse model and to elucidate the biological functionof Fancm in mouse development. Methods Fancm -/ - mice were generated by the CRISPR/ Cas9 method. We analyzedtheir body weight, birth rate, gender ratio, and fertility. Blood samples were analyzed by a complete blood count (CBC).Testes from the Fancm -/ - and wildtype littermates were subjected to pathophysiological assessment. Results C57BL/6background Fancm knockout mice were generated by ATG region deletion using the CRISPR/ Cas9 method. Western blottingindicated the complete depletion of FANCM protein in the testis lysate from Fancm -/ - mice. The Fancm -/ - mice showed nosign of embryonic lethality. However, there were significantly less female Fancm -/ - mice compared with male Fancm -/ - mice.There was no significant difference in body weight between the Fancm -/ - mice and wild type littermates. The CBC indicateda slight, but significant, increase in the Hb level. The Fancm -/ - mice were infertile. The male Fancm -/ - mice had a reducedtesticular size and abnormal testicular architecture. The IHC and TUNEL assays indicated increases in cell cycle arrest andapoptosis in spermatogenic cells. Conclusions Fancm gene knockout causes defects of male reproductive organ development in mice.
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