title: CRISPR/Cas9-mediated Dax1 knockout in the monkey recapitulates human AHC-HH.
abstract: Mutations in the DAX1 locus cause X-linked adrenal hypoplasia congenita (AHC) and hypogonadotropic hypogonadism (HH), which manifest with primary adrenal insufficiency and incomplete or absent sexual maturation, respectively. The associated defects in spermatogenesis can range from spermatogenic arrest to Sertoli cell only syndrome. Conclusions from Dax1 knockout mouse models provide only limited insight into AHC/HH disease mechanisms, because mouse models exhibit more extensive abnormalities in testicular development, including disorganized and incompletely formed testis cords with decreased number of peritubular myoid cells and male-to-female sex reversal. We previously reported successful clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated genome targeting in cynomolgus monkeys. Here, we describe a male fetal monkey in which targeted genome editing using CRISPR/Cas9 produced Dax1-null mutations in most somatic tissues and in the gonads. This DAX1-deficient monkey displayed defects in adrenal gland development and abnormal testis architecture with small cords, expanded blood vessels and extensive fibrosis. Sertoli cell formation was not affected. This phenotype strongly resembles findings in human patients with AHC-HH caused by mutations in DAX1. We further detected upregulation of Wnt/β-catenin-VEGF signaling in the fetal Dax1-deficient testis, suggesting abnormal activation of signaling pathways in the absence of DAX1 as one mechanism of AHC-HH. Our study reveals novel insight into the role of DAX1 in HH and provides proof-of-principle for the generation of monkey models of human disease via CRISPR/Cas9-mediated gene targeting.
authors.affiliation: Faculty of Life Science and Technology, Institute of Primate Translational Medicine Research, Kunming University of Science & Technology, Kunming 650500, China, Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China, National Engineering Research Center of Biomedicine and Animal Science, Kunming 650500, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Kang
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Shen
authors.affiliation: Faculty of Life Science and Technology, Institute of Primate Translational Medicine Research, Kunming University of Science & Technology, Kunming 650500, China, Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China, National Engineering Research Center of Biomedicine and Animal Science, Kunming 650500, China.
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Jianying
authors.initials: J
authors.lastname: Wang
authors.affiliation: Faculty of Life Science and Technology, Institute of Primate Translational Medicine Research, Kunming University of Science & Technology, Kunming 650500, China, Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China, National Engineering Research Center of Biomedicine and Animal Science, Kunming 650500, China.
authors.forename: Yuyu
authors.initials: Y
authors.lastname: Niu
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Yiqiang
authors.initials: Y
authors.lastname: Cui
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, National Resource Center for Mutant Mice, Nanjing 210061, China, School of Life Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, Shanghai 201210, China and.
authors.forename: Jiankui
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Faculty of Life Science and Technology, Institute of Primate Translational Medicine Research, Kunming University of Science & Technology, Kunming 650500, China, Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China, National Engineering Research Center of Biomedicine and Animal Science, Kunming 650500, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Xuejiang
authors.initials: X
authors.lastname: Guo
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, National Resource Center for Mutant Mice, Nanjing 210061, China, School of Life Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, Shanghai 201210, China and.
authors.forename: Bian
authors.initials: B
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China, xingxuhuang@mail.nju.edu.cn wji@kbimed.com shajh@njmu.edu.cn.
authors.forename: Jiahao
authors.initials: J
authors.lastname: Sha
authors.affiliation: Faculty of Life Science and Technology, Institute of Primate Translational Medicine Research, Kunming University of Science & Technology, Kunming 650500, China, Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China, National Engineering Research Center of Biomedicine and Animal Science, Kunming 650500, China, xingxuhuang@mail.nju.edu.cn wji@kbimed.com shajh@njmu.edu.cn.
authors.forename: Weizhi
authors.initials: W
authors.lastname: Ji
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, National Resource Center for Mutant Mice, Nanjing 210061, China, School of Life Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, Shanghai 201210, China and xingxuhuang@mail.nju.edu.cn wji@kbimed.com shajh@njmu.edu.cn.
authors.forename: Xingxu
authors.initials: X
authors.lastname: Huang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D064130:CRISPR-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D057137:DAX-1 Orphan Nuclear Receptor / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D000882:Haplorhini; D006801:Humans; D007006:Hypogonadism / Q000235:genetics / Q000378:metabolism; D008297:Male; D012708:Sertoli Cells / Q000378:metabolism; D013737:Testis / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D064130:CRISPR-Associated Proteins; D057137:DAX-1 Orphan Nuclear Receptor; D004268:DNA-Binding Proteins; D014157:Transcription Factors