In this work, we describe a single piggyBac transposon system containing both a tet-activator and a doxycycline-inducible expression cassette.We demonstrate that a gene product can be conditionally expressed from the integrated transposon and a second gene can be simultaneously targeted by a short hairpin Hair Removal RNA contained within the transposon, both in vivo and in mammalian and avian cell lines.We applied this system to stably modify chicken primordial germ cell (PGC) lines in vitro and induce a reporter gene at specific developmental stages after injection of the transposon-modified germ cells into chicken embryos.
We used this vector to express a constitutively-active AKT molecule during PGC migration to the forming gonad.We found that PGC migration was retarded and cells could not colonise the forming gonad.Correct levels Mirror Relocation of AKT activation are thus essential for germ cell migration during early embryonic development.