Site-specific integrase-mediated transgenesis in mice via pronuclear injection.
Proc Natl Acad Sci U S A
; 108(19): 7902-7, 2011 May 10.
Article
in En
| MEDLINE
| ID: mdl-21464299
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.
Full text:
1
Database:
MEDLINE
Main subject:
Mice, Transgenic
/
Gene Transfer Techniques
/
Integrases
Limits:
Animals
/
Pregnancy
Language:
En
Journal:
Proc Natl Acad Sci U S A
Year:
2011
Type:
Article
Affiliation country:
United States