title: CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein.
abstract: Previous works using human tripronuclear zygotes suggested that the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system could be a tool in correcting disease-causing mutations. However, whether this system was applicable in normal human (dual pronuclear, 2PN) zygotes was unclear. Here we demonstrate that CRISPR/Cas9 is also effective as a gene-editing tool in human 2PN zygotes. By injection of Cas9 protein complexed with the appropriate sgRNAs and homology donors into one-cell human embryos, we demonstrated efficient homologous recombination-mediated correction of point mutations in HBB and G6PD. However, our results also reveal limitations of this correction procedure and highlight the need for further research.
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, 100850, China. 1212qq-@163.com.|National Center for Protein Sciences Beijing, Life Sciences Park, Beijing, 102206, China. 1212qq-@163.com.
authors.forename: Lichun
authors.identifier: http://orcid.org/0000-0002-1636-9705
authors.initials: L
authors.lastname: Tang
authors.affiliation: Center for Reproductive Medicine, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510150, China.
authors.forename: Yanting
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: Center for Reproductive Medicine, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510150, China.
authors.forename: Hongzi
authors.initials: H
authors.lastname: Du
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, 100850, China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Gong
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, 100850, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Peng
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, 100850, China.
authors.forename: Buxi
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Center for Reproductive Medicine, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510150, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Lei
authors.affiliation: National Center for International Research of Biological Targeting Diagnosis and Therapy, Collaborative Innovation Center for Targeting Tumor Diagnosis and Therapy, Guangxi Medical University, Guangxi, 530021, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Zhao
authors.affiliation: Houston Fertility Institute, Houston, TX, 77063, USA.
authors.forename: Weihua
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Cardiology, Bayi Hospital Affiliated Nanjing University of Chineses Medicine, Nanjing, 210002, China. lxw7402@126.com.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Li
authors.affiliation: Center for Reproductive Medicine, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510150, China. ljq88gz@163.com.
authors.forename: Jianqiao
authors.initials: J
authors.lastname: Liu
mesh_terms: D001483:Base Sequence; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics; D000072669:Gene Editing / Q000379:methods*; D030342:Genetic Diseases, Inborn / Q000235:genetics / Q000628:therapy*; D015894:Genome, Human / Q000235:genetics; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D000072380:NIMA-Related Kinase 1 / Q000235:genetics*; D015053:Zygote / Q000254:growth & development*
chemical_list: D018398:Homeodomain Proteins; C064658:RAG-1 protein; D005954:Glucosephosphate Dehydrogenase; C000606914:NEK1 protein, human; D000072380:NIMA-Related Kinase 1; D004720:Endonucleases
keywords: CRISPR/Cas9; Cas9 protein; Gene modification; Homology-directed repair (HDR); Human zygotes