A new method for producing transgenic birds via direct in vivo transfection of primordial germ cells

Scott G. Tyack, Kristie A. Jenkins, Terri E O'Neil, Terry G Wise, Kirsten R. Morris, Matthew P Bruce, Scott McLeod, Alexander J. Wade, James McKay, Robert J. Moore, Karel A Schat, John W Lowenthal, Timothy J. Doran

Research output: Contribution to journalArticleResearchpeer-review

67 Citations (Scopus)

Abstract

Traditional methods of avian transgenesis involve complex manipulations involving either retroviral infection of blastoderms or the ex vivo manipulation of primordial germ cells (PGCs) followed by injection of the cells back into a recipient embryo. Unlike in mammalian systems, avian embryonic PGCs undergo a migration through the vasculature on their path to the gonad where they become the sperm or ova producing cells. In a development which simplifies the procedure of creating transgenic chickens we have shown that PGCs are directly transfectable in vivo using commonly available transfection reagents. We used Lipofectamine 2000 complexed with Tol2 transposon and transposase plasmids to stably transform PGCs in vivo generating transgenic offspring that express a reporter gene carried in the transposon. The process has been shown to be highly effective and as robust as the other methods used to create germ-line transgenic chickens while substantially reducing time, infrastructure and reagents required. The method described here defines a simple direct approach for transgenic chicken production, allowing researchers without extensive PGC culturing facilities or skills with retroviruses to produce transgenic chickens for wide-ranging applications in research, biotechnology and agriculture.

Original languageEnglish
Pages (from-to)1257-1264
Number of pages8
JournalTransgenic Research
Volume22
Issue number6
DOIs
Publication statusPublished - Dec 2013
Externally publishedYes

Keywords

  • Avian
  • Primordial germ cells
  • Transgenisis

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