Efficient TALEN-mediated gene targeting of chicken primordial germ cells [TECHNIQUES AND RESOURCES ARTICLE]

Lorna Taylor, Daniel F. Carlson, Sunil Nandi, Adrian Sherman, Scott C. Fahrenkrug, and Michael J. McGrew

In this work we use TALE nucleases (TALENs) to target a reporter construct to the DDX4 (vasa) locus in chicken primordial germ cells. Vasa is a key germ cell determinant in many animal species and is posited to control avian germ cell formation. We show that TALENs mediate homology directed repair of the DDX4 locus on the Z sex chromosome at high (8.1%) efficiencies. Large genetic deletions of 30kb encompassing the entire DDX4 locus were also created using a single TALEN pair. The targeted PGCs were germ line competent and were used to produce DDX4 null offspring. In DDX4 knockout chickens, PGCs are initially formed but are lost during meiosis in the developing ovary leading to adult female sterility. TALEN-mediated gene targeting in avian primordial germ cells is therefore an efficient process.

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