Germline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.

Genetic engineering can expand the utility of pigs for modeling human diseases, and for developing advanced therapeutic approaches. However, the inefficient production of transgenic pigs represents a technological bottleneck. Here, we assessed the hyperactive Sleeping Beauty (SB100X) transposon syst...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Wiebke Garrels, Lajos Mátés, Stephanie Holler, Anna Dalda, Ulrike Taylor, Björn Petersen, Heiner Niemann, Zsuzsanna Izsvák, Zoltán Ivics, Wilfried A Kues
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1120e48e7ded4acca0a4dd3bbfb2eeb1
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:1120e48e7ded4acca0a4dd3bbfb2eeb1
record_format	dspace
spelling	oai:doaj.org-article:1120e48e7ded4acca0a4dd3bbfb2eeb12021-11-18T06:47:08ZGermline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.1932-620310.1371/journal.pone.0023573https://doaj.org/article/1120e48e7ded4acca0a4dd3bbfb2eeb12011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21897845/?tool=EBIhttps://doaj.org/toc/1932-6203Genetic engineering can expand the utility of pigs for modeling human diseases, and for developing advanced therapeutic approaches. However, the inefficient production of transgenic pigs represents a technological bottleneck. Here, we assessed the hyperactive Sleeping Beauty (SB100X) transposon system for enzyme-catalyzed transgene integration into the embryonic porcine genome. The components of the transposon vector system were microinjected as circular plasmids into the cytoplasm of porcine zygotes, resulting in high frequencies of transgenic fetuses and piglets. The transgenic animals showed normal development and persistent reporter gene expression for >12 months. Molecular hallmarks of transposition were confirmed by analysis of 25 genomic insertion sites. We demonstrate germ-line transmission, segregation of individual transposons, and continued, copy number-dependent transgene expression in F1-offspring. In addition, we demonstrate target-selected gene insertion into transposon-tagged genomic loci by Cre-loxP-based cassette exchange in somatic cells followed by nuclear transfer. Transposase-catalyzed transgenesis in a large mammalian species expands the arsenal of transgenic technologies for use in domestic animals and will facilitate the development of large animal models for human diseases.Wiebke GarrelsLajos MátésStephanie HollerAnna DaldaUlrike TaylorBjörn PetersenHeiner NiemannZsuzsanna IzsvákZoltán IvicsWilfried A KuesPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 8, p e23573 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Wiebke Garrels Lajos Mátés Stephanie Holler Anna Dalda Ulrike Taylor Björn Petersen Heiner Niemann Zsuzsanna Izsvák Zoltán Ivics Wilfried A Kues Germline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.
description	Genetic engineering can expand the utility of pigs for modeling human diseases, and for developing advanced therapeutic approaches. However, the inefficient production of transgenic pigs represents a technological bottleneck. Here, we assessed the hyperactive Sleeping Beauty (SB100X) transposon system for enzyme-catalyzed transgene integration into the embryonic porcine genome. The components of the transposon vector system were microinjected as circular plasmids into the cytoplasm of porcine zygotes, resulting in high frequencies of transgenic fetuses and piglets. The transgenic animals showed normal development and persistent reporter gene expression for >12 months. Molecular hallmarks of transposition were confirmed by analysis of 25 genomic insertion sites. We demonstrate germ-line transmission, segregation of individual transposons, and continued, copy number-dependent transgene expression in F1-offspring. In addition, we demonstrate target-selected gene insertion into transposon-tagged genomic loci by Cre-loxP-based cassette exchange in somatic cells followed by nuclear transfer. Transposase-catalyzed transgenesis in a large mammalian species expands the arsenal of transgenic technologies for use in domestic animals and will facilitate the development of large animal models for human diseases.
format	article
author	Wiebke Garrels Lajos Mátés Stephanie Holler Anna Dalda Ulrike Taylor Björn Petersen Heiner Niemann Zsuzsanna Izsvák Zoltán Ivics Wilfried A Kues
author_facet	Wiebke Garrels Lajos Mátés Stephanie Holler Anna Dalda Ulrike Taylor Björn Petersen Heiner Niemann Zsuzsanna Izsvák Zoltán Ivics Wilfried A Kues
author_sort	Wiebke Garrels
title	Germline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.
title_short	Germline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.
title_full	Germline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.
title_fullStr	Germline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.
title_full_unstemmed	Germline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.
title_sort	germline transgenic pigs by sleeping beauty transposition in porcine zygotes and targeted integration in the pig genome.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/1120e48e7ded4acca0a4dd3bbfb2eeb1
work_keys_str_mv	AT wiebkegarrels germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT lajosmates germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT stephanieholler germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT annadalda germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT ulriketaylor germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT bjornpetersen germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT heinerniemann germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT zsuzsannaizsvak germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT zoltanivics germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome AT wilfriedakues germlinetransgenicpigsbysleepingbeautytranspositioninporcinezygotesandtargetedintegrationinthepiggenome
_version_	1718424398527987712

Germline transgenic pigs by Sleeping Beauty transposition in porcine zygotes and targeted integration in the pig genome.

Ejemplares similares