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题名: Gene transfer and mutagenesis mediated by Sleeping Beauty transposon in Nile tilapia (Oreochromis niloticus)
作者: He, Xiaozhen1, 2; Li, Jie1, 2; Long, Yong1; Song, Guili1; Zhou, Peiyong3; Liu, Qiuxiang3; Zhu, Zuoyan1; Cui, Zongbin1
通讯作者: Cui, ZB (reprint author), Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, 7 Donghu Rd, Wuhan 430072, Hubei, Peoples R China.
关键词: Sleeping Beauty ; Transposon ; Nile tilapia ; Gene transfer ; Mutagenesis
刊名: TRANSGENIC RESEARCH
发表日期: 2013-10-01
DOI: 10.1007/s11248-013-9693-8
卷: 22, 期:5, 页:913-924
收录类别: SCI
文章类型: Article
部门归属: [He, Xiaozhen ; Li, Jie ; Long, Yong ; Song, Guili ; Zhu, Zuoyan ; Cui, Zongbin] Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China ; [He, Xiaozhen ; Li, Jie] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China ; [Zhou, Peiyong ; Liu, Qiuxiang] Qingdao Tilapia Seed Multiplicat Farm, Jiaozhou 266317, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
资助者: National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892]
类目[WOS]: Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
摘要: The success of gene transfer has been demonstrated in many of vertebrate species, whereas the efficiency of producing transgenic animals remains pretty low due to the random integration of foreign genes into a recipient genome. The Sleeping Beauty (SB) transposon is able to improve the efficiency of gene transfer in zebrafish and mouse, but its activity in tilapia (Oreochromis niloticus) has yet to be characterized. Herein, we demonstrate the potential of using the SB transposon system as an effective tool for gene transfer and insertional mutagenesis in tilapia. A transgenic construct pT2/tiHsp70-SB11 was generated by subcloning the promoter of tilapia heat shock protein 70 (tiHsp70) gene, the SB11 transposase gene and the carp beta-actin gene polyadenylation signal into the second generation of SB transposon. Transgenic tilapia was produced by microinjection of this construct with in vitro synthesized capped SB11 mRNA. SB11 transposon was detected in 28.89 % of founders, 12.9 % of F1 and 43.75 % of F2. Analysis of genomic sequences flanking integrated transposons indicates that this transgenic tilapia line carries two copies of SB transposon, which landed into two different endogenous genes. Induced expression of SB11 gene after heat shock was detected using reverse transcription PCR in F2 transgenic individuals. In addition, the Cre/loxP system was introduced to delete the SB11 cassette for stabilization of gene interruption and bio-safety. These findings suggest that the SB transposon system is active and can be used for efficient gene transfer and insertional mutagenesis in tilapia.
英文摘要: The success of gene transfer has been demonstrated in many of vertebrate species, whereas the efficiency of producing transgenic animals remains pretty low due to the random integration of foreign genes into a recipient genome. The Sleeping Beauty (SB) transposon is able to improve the efficiency of gene transfer in zebrafish and mouse, but its activity in tilapia (Oreochromis niloticus) has yet to be characterized. Herein, we demonstrate the potential of using the SB transposon system as an effective tool for gene transfer and insertional mutagenesis in tilapia. A transgenic construct pT2/tiHsp70-SB11 was generated by subcloning the promoter of tilapia heat shock protein 70 (tiHsp70) gene, the SB11 transposase gene and the carp beta-actin gene polyadenylation signal into the second generation of SB transposon. Transgenic tilapia was produced by microinjection of this construct with in vitro synthesized capped SB11 mRNA. SB11 transposon was detected in 28.89 % of founders, 12.9 % of F1 and 43.75 % of F2. Analysis of genomic sequences flanking integrated transposons indicates that this transgenic tilapia line carries two copies of SB transposon, which landed into two different endogenous genes. Induced expression of SB11 gene after heat shock was detected using reverse transcription PCR in F2 transgenic individuals. In addition, the Cre/loxP system was introduced to delete the SB11 cassette for stabilization of gene interruption and bio-safety. These findings suggest that the SB transposon system is active and can be used for efficient gene transfer and insertional mutagenesis in tilapia.
关键词[WOS]: TRANSGENIC TILAPIA ; GROWTH-HORMONE ; ZEBRAFISH EMBRYOS ; COMMON CARP ; EXPRESSION ; DNA ; FISH ; CELLS ; PROMOTER ; VECTORS
语种: 英语
项目资助者: National High-tech RD (863) Program(2007AA10Z164) ; National Natural Science Foundation of China(31101892)
WOS记录号: WOS:000324800000004
ISSN号: 0962-8819
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/19608
Appears in Collections:水生生物分子与细胞生物学研究中心_期刊论文

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作者单位: 1.Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
3.Qingdao Tilapia Seed Multiplicat Farm, Jiaozhou 266317, Peoples R China

Recommended Citation:
He, Xiaozhen; Li, Jie; Long, Yong; Song, Guili; Zhou, Peiyong; Liu, Qiuxiang; Zhu, Zuoyan; Cui, Zongbin.Gene transfer and mutagenesis mediated by Sleeping Beauty transposon in Nile tilapia (Oreochromis niloticus),TRANSGENIC RESEARCH,2013,22(5):913-924
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