中国科学院水生生物研究所机构知识库
Advanced  
IHB OpenIR  > 水生生物分子与细胞生物学研究中心  > 期刊论文
题名: Effective Gene Trapping Mediated by Sleeping Beauty Transposon
作者: Song, Guili1, 2; Li, Qing1; Long, Yong1; Gu, Qilin1, 2; Hackett, Perry B.3; Cui, Zongbin1
通讯作者: Song, GL (reprint author), Chinese Acad Sci, Inst Hydrobiol, Key Lab Aquat Biodivers & Conservat, Wuhan, Hubei, Peoples R China.
关键词: EMBRYONIC STEM-CELLS ; DEVELOPMENTALLY-REGULATED GENES ; MESSENGER-RNA DECAY ; INSERTIONAL MUTAGENESIS ; HEPATOCELLULAR-CARCINOMA ; FUNCTIONAL GENOMICS ; ENU MUTAGENESIS ; LARGE-SCALE ; ZEBRAFISH ; MOUSE
刊名: PLOS ONE
发表日期: 2012-08-31
DOI: 10.1371/journal.pone.0044123
卷: 7, 期:8, 页:e44123
收录类别: SCI
文章类型: Article
部门归属: [Song, Guili; Li, Qing; Long, Yong; Gu, Qilin; Cui, Zongbin] Chinese Acad Sci, Inst Hydrobiol, Key Lab Aquat Biodivers & Conservat, Wuhan, Hubei, Peoples R China; [Song, Guili; Gu, Qilin] Chinese Acad Sci, Grad Univ, Beijing, Peoples R China; [Hackett, Perry B.] Univ Minnesota, Dept Genet Cell Biol & Dev, Minneapolis, MN USA
WOS标题词: Science & Technology
资助者: National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442]
类目[WOS]: Multidisciplinary Sciences
研究领域[WOS]: Science & Technology - Other Topics
摘要: Gene trapping is a high-throughput approach to elucidate gene functions by disrupting and recapitulating expression of genes in a target genome. A number of transposon-based gene-trapping systems are developed for mutagenesis in cells and model organisms, but there is still much room for the improvement of their efficiency in gene disruption and mutation. Herein, a gene-trapping system mediated by Sleeping Beauty (SB) transposon was developed by inclusion of three functional cassettes. The mutation cassette can abrogate the splice of trapped genes and terminate their translation. Once an endogenous gene is captured, the finding cassette independently drives the translation of reporter gene in HeLa cells and zebrafish embryos. The efficiency cassette controls the remobilization of integrated traps through inducible expression of SB gene. Analysis of transposon-genome junctions indicate that most of trap cassettes are integrated into an intron without an obvious 39 bias. The transcription of trapped genes was abrogated by alternative splicing of the mutation cassette. In addition, integrated transposons can be induced to excise from their original insertion sites. Furthermore, the Cre/LoxP system was introduced to delete the efficiency cassette for stabilization of gene interruption and bio-safety. Thus, this gene-trap vector is an alternative and effective tool for the capture and disruption of endogenous genes in vitro and in vivo.
英文摘要: Gene trapping is a high-throughput approach to elucidate gene functions by disrupting and recapitulating expression of genes in a target genome. A number of transposon-based gene-trapping systems are developed for mutagenesis in cells and model organisms, but there is still much room for the improvement of their efficiency in gene disruption and mutation. Herein, a gene-trapping system mediated by Sleeping Beauty (SB) transposon was developed by inclusion of three functional cassettes. The mutation cassette can abrogate the splice of trapped genes and terminate their translation. Once an endogenous gene is captured, the finding cassette independently drives the translation of reporter gene in HeLa cells and zebrafish embryos. The efficiency cassette controls the remobilization of integrated traps through inducible expression of SB gene. Analysis of transposon-genome junctions indicate that most of trap cassettes are integrated into an intron without an obvious 39 bias. The transcription of trapped genes was abrogated by alternative splicing of the mutation cassette. In addition, integrated transposons can be induced to excise from their original insertion sites. Furthermore, the Cre/LoxP system was introduced to delete the efficiency cassette for stabilization of gene interruption and bio-safety. Thus, this gene-trap vector is an alternative and effective tool for the capture and disruption of endogenous genes in vitro and in vivo.
关键词[WOS]: EMBRYONIC STEM-CELLS ; DEVELOPMENTALLY-REGULATED GENES ; MESSENGER-RNA DECAY ; INSERTIONAL MUTAGENESIS ; HEPATOCELLULAR-CARCINOMA ; FUNCTIONAL GENOMICS ; ENU MUTAGENESIS ; LARGE-SCALE ; ZEBRAFISH ; MOUSE
语种: 英语
WOS记录号: WOS:000308221300053
ISSN号: 1932-6203
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/17268
Appears in Collections:水生生物分子与细胞生物学研究中心_期刊论文

Files in This Item: Download All
File Name/ File Size Content Type Version Access License
Effective Gene Trapping Mediated by Sleeping Beauty Transposon.pdf(2121KB)----开放获取--View Download

作者单位: 1.Chinese Acad Sci, Inst Hydrobiol, Key Lab Aquat Biodivers & Conservat, Wuhan, Hubei, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China
3.Univ Minnesota, Dept Genet Cell Biol & Dev, Minneapolis, MN USA

Recommended Citation:
Song, Guili; Li, Qing; Long, Yong; Gu, Qilin; Hackett, Perry B.; Cui, Zongbin.Effective Gene Trapping Mediated by Sleeping Beauty Transposon,PLOS ONE,2012,7(8):e44123
Service
Recommend this item
Sava as my favorate item
Show this item's statistics
Export Endnote File
Google Scholar
Similar articles in Google Scholar
[Song, Guili]'s Articles
[Li, Qing]'s Articles
[Long, Yong]'s Articles
CSDL cross search
Similar articles in CSDL Cross Search
[Song, Guili]‘s Articles
[Li, Qing]‘s Articles
[Long, Yong]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
Add to CiteULike Add to Connotea Add to Del.icio.us Add to Digg Add to Reddit
文件名: Effective Gene Trapping Mediated by Sleeping Beauty Transposon.pdf
格式: Adobe PDF
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Copyright © 2007-2016  中国科学院水生生物研究所 - Feedback
Powered by CSpace