中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
首页 机构 成果 学者
  
  1. CAS IR Grid
  2. 上海营养与健康研究所
  3. 中国科学院上海生命科学研究院营养科学研究所
In Vivo Exon Replacement in the Mouse Atp7b Gene by the Cas9 System

文献类型：期刊论文

作者Liu, Lili2; Chang, Qiurong2; Xing, Fengying2; Yan, Guofeng2; Fu, Li2; Wang, Huiyang2; Ma, Zhengwen2; Chen, Xuejin2; Li, Yao2; Cao, Jianchang1
刊名HUMAN GENE THERAPY
出版日期2019
卷号30期号:9页码:1079-1092
关键词exon replacement CRISPR Cas9 rAAV ATP7B
ISSN号1043-0342
DOI10.1089/hum.2019.037
文献子类Article
英文摘要The application of CRISPR/Cas9 has opened a new era in gene therapy, making it possible to correct mutated genomes in vivo. Exon replacement can correct many mutations and has potential clinical value. In this study, we used a lentivirus-delivered transgene to obtain transgenic mice in which Cas9 and green fluorescent protein (GFP) were driven by the hTBG promoter and were specifically expressed in the liver. In Cas9-positive mice, only similar to 11.6% of hepatocytes were GFP positive. The newborn Cas9-positive F1 mice were injected via the temporal vein with rAAV carrying a modified homologous replacement sequence for exon 8 of Atp7b and a pair of single-strand guide RNAs targeting the introns surrounding exon 8. When the Cas9-positive hepatocytes were sorted and analyzed by PCR and next-generation deep sequencing with different labels, similar to 16.34 +/- 4.02% to 19.37 +/- 6.50% of the analyzed copies of exon 8 were replaced by the donor template in the genome of GFP-positive hepatocytes, that is, 1.81 +/- 0.29% to 2.09 +/- 0.54% replacement occurred in all liver genomes. However, when rAAV carrying a modified homologous replacement sequence was injected into the adult spCas9 mice, a double-cut deletion ratio of up to 99%, only about 1.10-1.13% of the exon 8 replacement rate was detected in Cas9-positive hepatocytes. This study is the first to achieve exon replacement via CRISPR/Cas9, which will benefit research on CRISPR/Cas9 technology for gene therapy.
学科主题Biotechnology & Applied Microbiology ; Genetics & Heredity ; Research & Experimental Medicine
WOS关键词WILSON-DISEASE MUTATIONS ; BINDING GLOBULIN GENE ; LIVER ; DNA ; EXPRESSION ; EFFICIENT ; THERAPY ; REPEATS ; MODEL ; MICE
语种英语
WOS记录号WOS:000474172600001
出版者MARY ANN LIEBERT, INC
版本出版稿
源URL[http://202.127.25.144/handle/331004/547]  
专题中国科学院上海生命科学研究院营养科学研究所
作者单位1.Univ Chinese Acad Sci, Chinese Acad Sci, CAS Key Lab Tissue Microenvironm & Tumor, Shanghai Inst Nutr & Hlth,Shanghai Inst Biol Sci, Shanghai, Peoples R China;
2.Shanghai Jiao Tong Univ, Sch Med, Dept Lab Anim Sci, Shanghai, Peoples R China;
3.Kunming Univ Sci & Technol, Inst Primate Translat Med, Yunnan Key Lab Primate Biomed Res, Kunming, Yunnan, Peoples R China,
推荐引用方式
GB/T 7714		 Liu, Lili,Chang, Qiurong,Xing, Fengying,et al. In Vivo Exon Replacement in the Mouse Atp7b Gene by the Cas9 System[J]. HUMAN GENE THERAPY,2019,30(9):1079-1092.
APA Liu, Lili.,Chang, Qiurong.,Xing, Fengying.,Yan, Guofeng.,Fu, Li.,...&,.(2019).In Vivo Exon Replacement in the Mouse Atp7b Gene by the Cas9 System.HUMAN GENE THERAPY,30(9),1079-1092.
MLA Liu, Lili,et al."In Vivo Exon Replacement in the Mouse Atp7b Gene by the Cas9 System".HUMAN GENE THERAPY 30.9(2019):1079-1092.

入库方式： OAI收割

来源：上海营养与健康研究所

浏览0
下载0
收藏0
其他版本

除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。