2015 Fiscal Year Final Research Report
Development and verification of the next generation transgenesis tools
| Project/Area Number |
25290035
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Partial Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Tokai University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
MOCHIDA Keiji 理化学研究所, バイオリソースセンター, 専任技師 (60312287)
SHITARA Hiroshi 東京都医学総合研究所, 基盤技術研究センター, 主任基盤技術研究職員 (90321885)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Keywords | トランスジェニックマウス / PITT / Cre-loxP / PhiC31 / Rosa26 |
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| Outline of Final Research Achievements |
We recently developed technology termed “pronuclear injection (PI)-based Targeted Transgenesis (PITT)” that allows targeted insertion of single-copy transgene into the predetermined locus through PI. PITT method enables to generate Tg mice showing reliable transgene expression pattern. Here, we developed an improved-PITT (i-PITT) system by combining Cre-loxP and PhiC31-attP/B systems directly under C57BL/6N inbred strain. The targeted Tg efficiency in the i-PITT was improved up to 67%, which is by-far the best Tg rate reported. Furthermore, the system could generate multiple Tg mice simultaneously: injection of up to three different Tg cassettes in a single injection session into as less as 181 zygotes resulted in production of all three separate Tg DNA containing targeted Tg mice. Standard protocol for PITT was published and the seed mouse strain is available from RIKEN BRC.
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| Free Research Field |
マウス遺伝子工学
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