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Mol Genet Genomics. 2017 Jun;292(3):525-533. doi: 10.1007/s00438-017-1299-z. Epub 2017 Mar 1.

CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein.

Author information

1
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, 100850, China. 1212qq-@163.com.
2
National Center for Protein Sciences Beijing, Life Sciences Park, Beijing, 102206, China. 1212qq-@163.com.
3
Center for Reproductive Medicine, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510150, China.
4
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, 100850, China.
5
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.
6
Houston Fertility Institute, Houston, TX, 77063, USA.
7
Department of Cardiology, Bayi Hospital Affiliated Nanjing University of Chineses Medicine, Nanjing, 210002, China. lxw7402@126.com.
8
Center for Reproductive Medicine, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510150, China. ljq88gz@163.com.

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.

KEYWORDS:

CRISPR/Cas9; Cas9 protein; Gene modification; Homology-directed repair (HDR); Human zygotes

PMID:
28251317
DOI:
10.1007/s00438-017-1299-z
[Indexed for MEDLINE]

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