IHB OpenIR  > 水生生物分子与细胞生物学研究中心  > 期刊论文
Gene transfer and mutagenesis mediated by Sleeping Beauty transposon in Nile tilapia (Oreochromis niloticus)
He, Xiaozhen1,2; Li, Jie1,2; Long, Yong1; Song, Guili1; Zhou, Peiyong3; Liu, Qiuxiang3; Zhu, Zuoyan1; Cui, Zongbin1; Cui, ZB (reprint author), Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, 7 Donghu Rd, Wuhan 430072, Hubei, Peoples R China.
2013-10-01
Source PublicationTRANSGENIC RESEARCH
ISSN0962-8819
Volume22Issue:5Pages:913-924
AbstractThe 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.; 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.
SubtypeArticle
KeywordSleeping Beauty Transposon Nile Tilapia Gene Transfer Mutagenesis
Department[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
DOI10.1007/s11248-013-9693-8
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationNational High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892] ; National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892] ; National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892] ; National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892]
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS IDWOS:000324800000004
WOS KeywordTRANSGENIC TILAPIA ; GROWTH-HORMONE ; ZEBRAFISH EMBRYOS ; COMMON CARP ; EXPRESSION ; DNA ; FISH ; CELLS ; PROMOTER ; VECTORS
Funding OrganizationNational High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892] ; National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892] ; National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892] ; National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892]
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/19608
Collection水生生物分子与细胞生物学研究中心_期刊论文
Corresponding AuthorCui, ZB (reprint author), Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, 7 Donghu Rd, Wuhan 430072, Hubei, Peoples R China.
Affiliation1.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
GB/T 7714
He, Xiaozhen,Li, Jie,Long, Yong,et al. Gene transfer and mutagenesis mediated by Sleeping Beauty transposon in Nile tilapia (Oreochromis niloticus)[J]. TRANSGENIC RESEARCH,2013,22(5):913-924.
APA He, Xiaozhen.,Li, Jie.,Long, Yong.,Song, Guili.,Zhou, Peiyong.,...&Cui, ZB .(2013).Gene transfer and mutagenesis mediated by Sleeping Beauty transposon in Nile tilapia (Oreochromis niloticus).TRANSGENIC RESEARCH,22(5),913-924.
MLA He, Xiaozhen,et al."Gene transfer and mutagenesis mediated by Sleeping Beauty transposon in Nile tilapia (Oreochromis niloticus)".TRANSGENIC RESEARCH 22.5(2013):913-924.
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