Journal
PLOS ONE
Volume 8, Issue 11, Pages -Publisher
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0077222
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Funding
- Biotechnology and Biological Sciences Research Council
- BBSRC [BB/H023593/1]
- Biotechnology and Biological Sciences Research Council [BBS/E/D/20320000, BBS/E/R/00001607, BBS/E/D/20221658, BB/H023593/1] Funding Source: researchfish
- BBSRC [BB/H023593/1, BBS/E/R/00001607, BBS/E/D/20320000, BBS/E/D/20221658] Funding Source: UKRI
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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 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 of AKT activation are thus essential for germ cell migration during early embryonic development.
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