Site specific integrase mediated transgenesis in mice via pronuclear injection

Tasic B, Hippenmeyer S, Wang C, Gamboa M, Zong H, Chen Tsai Y, Luo L. 2011. Site specific integrase mediated transgenesis in mice via pronuclear injection. PNAS. 108(19), 7902–7907.

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Journal Article | Published
Author
Tasic, Bosiljka; Hippenmeyer, SimonIST Austria ; Wang, Charlene; Gamboa, Matthew; Zong, Hui; Chen-Tsai, Yanru; Luo, Liqun
Abstract
Microinjection of recombinant DNA into zygotic pronuclei has been widely used for producing transgenic mice. However, with this method, the insertion site, integrity, and copy number of the transgene cannot be controlled. Here, we present an integrase-based approach to produce transgenic mice via pronuclear injection, whereby an intact single-copy transgene can be inserted into predetermined chromosomal loci with high efficiency (up to 40%), and faithfully transmitted through generations. We show that neighboring transgenic elements and bacterial DNA within the transgene cause profound silencing and expression variability of the transgenic marker. Removal of these undesirable elements leads to global high-level marker expression from transgenes driven by a ubiquitous promoter. We also obtained faithful marker expression from a tissue-specific promoter. The technique presented here will greatly facilitate murine transgenesis and precise structure/function dissection of mammalian gene function and regulation in vivo.
Publishing Year
Date Published
2011-05-10
Journal Title
PNAS
Volume
108
Issue
19
Page
7902 - 7907
IST-REx-ID

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Tasic B, Hippenmeyer S, Wang C, et al. Site specific integrase mediated transgenesis in mice via pronuclear injection. PNAS. 2011;108(19):7902-7907. doi:10.1073/pnas.1019507108
Tasic, B., Hippenmeyer, S., Wang, C., Gamboa, M., Zong, H., Chen Tsai, Y., & Luo, L. (2011). Site specific integrase mediated transgenesis in mice via pronuclear injection. PNAS. National Academy of Sciences. https://doi.org/10.1073/pnas.1019507108
Tasic, Bosiljka, Simon Hippenmeyer, Charlene Wang, Matthew Gamboa, Hui Zong, Yanru Chen Tsai, and Liqun Luo. “Site Specific Integrase Mediated Transgenesis in Mice via Pronuclear Injection.” PNAS. National Academy of Sciences, 2011. https://doi.org/10.1073/pnas.1019507108.
B. Tasic et al., “Site specific integrase mediated transgenesis in mice via pronuclear injection,” PNAS, vol. 108, no. 19. National Academy of Sciences, pp. 7902–7907, 2011.
Tasic B, Hippenmeyer S, Wang C, Gamboa M, Zong H, Chen Tsai Y, Luo L. 2011. Site specific integrase mediated transgenesis in mice via pronuclear injection. PNAS. 108(19), 7902–7907.
Tasic, Bosiljka, et al. “Site Specific Integrase Mediated Transgenesis in Mice via Pronuclear Injection.” PNAS, vol. 108, no. 19, National Academy of Sciences, 2011, pp. 7902–07, doi:10.1073/pnas.1019507108.

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