• Register
X
Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.

X

Leaving Community

Are you sure you want to leave this community? Leaving the community will revoke any permissions you have been granted in this community.

No
Yes

Site-specific integrase-mediated transgenesis in mice via pronuclear injection.

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.

Pubmed ID: 21464299

Authors

  • Tasic B
  • Hippenmeyer S
  • Wang C
  • Gamboa M
  • Zong H
  • Chen-Tsai Y
  • Luo L

Journal

Proceedings of the National Academy of Sciences of the United States of America

Publication Data

May 10, 2011

Associated Grants

  • Agency: NINDS NIH HHS, Id: R01 NS050835
  • Agency: NINDS NIH HHS, Id: R01-NS050835
  • Agency: Howard Hughes Medical Institute, Id:

Mesh Terms

  • Animals
  • Attachment Sites, Microbiological
  • Base Sequence
  • Binding Sites
  • DNA Primers
  • Female
  • Gene Expression
  • Gene Transfer Techniques
  • Genetic Markers
  • Green Fluorescent Proteins
  • Integrases
  • Male
  • Mice
  • Mice, Transgenic
  • Microinjections
  • Pregnancy
  • Recombinant Proteins
  • Tissue Distribution