IHB OpenIR  > 水生生物分子与细胞生物学研究中心  > 期刊论文
Effective Gene Trapping Mediated by Sleeping Beauty Transposon
Song, Guili1,2; Li, Qing1; Long, Yong1; Gu, Qilin1,2; Hackett, Perry B.3; Cui, Zongbin1; Song, GL (reprint author), Chinese Acad Sci, Inst Hydrobiol, Key Lab Aquat Biodivers & Conservat, Wuhan, Hubei, Peoples R China.
2012-08-31
Source PublicationPLOS ONE
ISSN1932-6203
Volume7Issue:8Pages:e44123
AbstractGene 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.; 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.
SubtypeArticle
KeywordEmbryonic Stem-cells Developmentally-regulated Genes Messenger-rna Decay Insertional Mutagenesis Hepatocellular-carcinoma Functional Genomics Enu Mutagenesis Large-scale Zebrafish Mouse
Department[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
DOI10.1371/journal.pone.0044123
WOS HeadingsScience & Technology
Funding OrganizationNational Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442]
Indexed BySCI
Language英语
WOS Research AreaScience & Technology - Other Topics
WOS SubjectMultidisciplinary Sciences
WOS IDWOS:000308221300053
WOS KeywordEMBRYONIC STEM-CELLS ; DEVELOPMENTALLY-REGULATED GENES ; MESSENGER-RNA DECAY ; INSERTIONAL MUTAGENESIS ; HEPATOCELLULAR-CARCINOMA ; FUNCTIONAL GENOMICS ; ENU MUTAGENESIS ; LARGE-SCALE ; ZEBRAFISH ; MOUSE
Funding OrganizationNational Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [30871442]
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/17268
Collection水生生物分子与细胞生物学研究中心_期刊论文
Corresponding AuthorSong, GL (reprint author), Chinese Acad Sci, Inst Hydrobiol, Key Lab Aquat Biodivers & Conservat, Wuhan, Hubei, Peoples R China.
Affiliation1.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
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..,...&Song, GL .(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.
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