Effective Gene Trapping Mediated by Sleeping Beauty Transposon

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	Effective Gene Trapping Mediated by Sleeping Beauty Transposon
	Song, Guili 1,2; Li, Qing 1; Long, Yong 1; Gu, Qilin 1,2; Hackett, Perry B.3; Cui, Zongbin 1
刊名	PLOS ONE
	2012-08-31
卷号	7 期号:8 页码:e44123
关键词	EMBRYONIC STEM-CELLS DEVELOPMENTALLY-REGULATED GENES MESSENGER-RNA DECAY INSERTIONAL MUTAGENESIS HEPATOCELLULAR-CARCINOMA FUNCTIONAL GENOMICS ENU MUTAGENESIS LARGE-SCALE ZEBRAFISH MOUSE
ISSN号	1932-6203
通讯作者	Song, GL (reprint author), Chinese Acad Sci, Inst Hydrobiol, Key Lab Aquat Biodivers & Conservat, Wuhan, Hubei, Peoples R China.
中文摘要	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.
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.
WOS标题词	Science & Technology
类目[WOS]	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
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
收录类别	SCI
资助信息	National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442]
语种	英语
WOS记录号	WOS:000308221300053
公开日期	2013-01-09
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/342005/17268]
专题	水生生物研究所_水生生物分子与细胞生物学研究中心_期刊论文
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
推荐引用方式 GB/T 7714	Song, Guili,Li, Qing,Long, Yong,et al. Effective Gene Trapping Mediated by Sleeping Beauty Transposon[J]. PLOS ONE,2012,7(8):e44123.
APA	Song, Guili,Li, Qing,Long, Yong,Gu, Qilin,Hackett, Perry B.,&Cui, Zongbin.(2012).Effective Gene Trapping Mediated by Sleeping Beauty Transposon.PLOS ONE,7(8),e44123.
MLA	Song, Guili,et al."Effective Gene Trapping Mediated by Sleeping Beauty Transposon".PLOS ONE 7.8(2012):e44123.