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CRISPR/Cas9-AAV Mediated Knock-in at NRL Locus in Human Embryonic Stem Cells
Xianglian Ge1; Haitao Xi2; Fayu Yang1; Xiao Zhi1; Yanghua Fu3; Ding Chen1; Ren-He Xu4; Ge Lin5; Jia Qu1; Junzhao Zhao3; Feng Gu1
2016
Source PublicationMolecular Therapy - Nucleic Acids
ISSN21622531
Volume5
Abstract

Clustered interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated genome engineering technologies are sparking a new revolution in biological research. This technology efficiently induces DNA double strand breaks at the targeted genomic sequence and results in indel mutations by the error-prone process of nonhomologous end joining DNA repair or homologous recombination with a DNA repair template. The efficiency of genome editing with CRISPR/Cas9 alone in human embryonic stem cells is still low. Gene targeting with adeno-associated virus (AAV) vectors has been demonstrated in multiple human cell types with maximal targeting frequencies without engineered nucleases. However, whether CRISPR/Cas9-mediated double strand breaks and AAV based donor DNA mediated homologous recombination approaches could be combined to create a novel CRISPR/Cas9-AAV genetic tool for highly specific gene editing is not clear. Here we demonstrate that using CRISPR/Cas9-AAV, we could successfully knock-in a DsRed reporter gene at the basic motifleucine zipper transcription factor (NRL) locus in human embryonic stem cells. For the first time, this study provides the proof of principle that these two technologies can be used together. CRISPR/Cas9-AAV, a new genome editing tool, offers a platform for the manipulation of human genome.

KeywordAav Crispr/cas9 Homologous Recombination Human Embryonic Stem Cells Next Generation Sequencing Nrl
DOI10.1038/mtna.2016.100
URLView the original
Language英语
WOS Research AreaResearch & Experimental Medicine
WOS SubjectMedicine, Research & Experimental
WOS IDWOS:000395791100003
PublisherCELL PRESS, 600 TECHNOLOGY SQUARE, 5TH FLOOR, CAMBRIDGE, MA 02139 USA
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被引频次[WOS]:5   [WOS记录]     [WOS相关记录]
Document TypeJournal article
专题University of Macau
Corresponding AuthorJunzhao Zhao; Feng Gu
Affiliation1.School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, State Key Laboratory Cultivation Base and Key Laboratory of Vision Science, Ministry of Health and Zhejiang Provincial Key Laboratory of Ophthalmology and Optometry, Wenzhou,
2.Reproductive medicine Center, The Second Affiliated Hospital, Wenzhou Medical University, Wenzhou, China
3.Reproductive medicine Center, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou, China
4.Faculty of Health Sciences, University of Macau, Macau, China
5.Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China. Correspondence: Feng Gu, Center for Vision Research, Eye Hospital, Wenzhou Medical University, 270 Xueyuan West Road, Wenzhou, Zhejiang 325027, China
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Xianglian Ge,Haitao Xi,Fayu Yang,et al. CRISPR/Cas9-AAV Mediated Knock-in at NRL Locus in Human Embryonic Stem Cells[J]. Molecular Therapy - Nucleic Acids,2016,5.
APA Xianglian Ge.,Haitao Xi.,Fayu Yang.,Xiao Zhi.,Yanghua Fu.,...&Feng Gu.(2016).CRISPR/Cas9-AAV Mediated Knock-in at NRL Locus in Human Embryonic Stem Cells.Molecular Therapy - Nucleic Acids,5.
MLA Xianglian Ge,et al."CRISPR/Cas9-AAV Mediated Knock-in at NRL Locus in Human Embryonic Stem Cells".Molecular Therapy - Nucleic Acids 5(2016).
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