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Using TARGATT™ Technology to Generate Site-Specific Transgenic Mice

  • Ruby Yanru Chen-TsaiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1874)

Abstract

The discovery of new gene editing tools in the past several years has moved the transgenic field to a new level. The traditional random transgenesis method by pronuclear microinjection has been largely replaced by targeted or site-specific transgenic technologies without the need of homologous recombination in embryonic stem (ES) cells. In this chapter, I describe detailed protocols of an integrase-based approach, trademarked as “TARGATT™” (target attP), to produce site-specific transgenic mice via pronuclear microinjection, whereby an intact single-copy transgene can be inserted into a predetermined chromosomal locus with high efficiency (up to 40%), and faithfully transmitted through generations. This system allows high-level global transgene expression or tissue-specific expression depending on the promoter used, or inducible expression such as induced by tetracycline or doxycycline. Using this approach, site-specific transgenic mice can be generated as fast as in 3 months. The technique presented here greatly facilitates murine transgenesis and precise structure/function dissection of mammalian gene function and regulation in vivo.

Key words

Pronuclear microinjection PhiC31 integrase TARGATT™ Site-specific transgenic Rosa26 knockin H11 locus attP attB 

Notes

Acknowledgment

The author would like to thank all Applied StemCell, Inc. employees who have contributed to improving the TARGATT™ technologies. Thanks also go to our customers who entrusted us on generating TARGATT™ models for their research.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Applied StemCell, Inc.MilpitasUSA

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