A novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.

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...

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Autores principales:	James D Glover, Lorna Taylor, Adrian Sherman, Caroline Zeiger-Poli, Helen M Sang, Michael J McGrew
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0eb05673365341ecaf68a58a5c7dffb9
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spelling	oai:doaj.org-article:0eb05673365341ecaf68a58a5c7dffb92021-11-18T08:48:38ZA novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.1932-620310.1371/journal.pone.0077222https://doaj.org/article/0eb05673365341ecaf68a58a5c7dffb92013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24223709/?tool=EBIhttps://doaj.org/toc/1932-6203In 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.James D GloverLorna TaylorAdrian ShermanCaroline Zeiger-PoliHelen M SangMichael J McGrewPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 11, p e77222 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q James D Glover Lorna Taylor Adrian Sherman Caroline Zeiger-Poli Helen M Sang Michael J McGrew A novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.
description	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.
format	article
author	James D Glover Lorna Taylor Adrian Sherman Caroline Zeiger-Poli Helen M Sang Michael J McGrew
author_facet	James D Glover Lorna Taylor Adrian Sherman Caroline Zeiger-Poli Helen M Sang Michael J McGrew
author_sort	James D Glover
title	A novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.
title_short	A novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.
title_full	A novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.
title_fullStr	A novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.
title_full_unstemmed	A novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.
title_sort	novel piggybac transposon inducible expression system identifies a role for akt signalling in primordial germ cell migration.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/0eb05673365341ecaf68a58a5c7dffb9
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A novel piggyBac transposon inducible expression system identifies a role for AKT signalling in primordial germ cell migration.

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