| 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 Publication | TRANSGENIC RESEARCH
 |
| ISSN | 0962-8819 |
| Volume | 22Issue:5Pages:913-924 |
| Abstract | 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.; 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. |
| Subtype | Article |
| Keyword | Sleeping 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 |
| DOI | 10.1007/s11248-013-9693-8 |
| WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
| Funding Organization | 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] ; National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892] |
| Indexed By | SCI |
| Language | 英语 |
| WOS Research Area | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| WOS Subject | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| WOS ID | WOS:000324800000004 |
| WOS Keyword | TRANSGENIC TILAPIA ; GROWTH-HORMONE ; ZEBRAFISH EMBRYOS ; COMMON CARP ; EXPRESSION ; DNA ; FISH ; CELLS ; PROMOTER ; VECTORS |
| Funding Organization | 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] ; National High-tech RD (863) Program [2007AA10Z164]; National Natural Science Foundation of China [31101892] |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://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 Author | Cui, ZB (reprint author), Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, 7 Donghu Rd, Wuhan 430072, Hubei, Peoples R China. |
| Affiliation | 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 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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