Genetic modification of primordial germ cells by gene trapping, gene targeting, and phiC31 integrase

Molecular Reproduction and Development
Philip A LeightonRobert J Etches

Abstract

The genome of germline committed cells is thought to be protected by mechanisms of transcriptional silencing, posing a barrier to transgenesis using cultured germline cells. We found that selection for transgene integration into the primordial germ cell genome required that the transgenes be flanked by the chicken beta-globin insulator. However, integration frequency was low, and sequencing of the insertion sites revealed that the transgenes preferentially inserted into active promoter regions, implying that silencing prohibited recovery of insertions in other regions. Much higher frequencies of integration were achieved when the phiC31 integrase was used to insert transgenes into endogenous pseudo attP sites. Despite the evidence for transcriptional silencing in PGCs, gene targeting of a nonexpressed gene was also achieved. The ability to make genetic modifications in PGCs provides unprecedented opportunities to study the biology of PGCs, as well as produce transgenic chickens for applications in biotechnology and developmental biology.

References

Mar 1, 1976·Developmental Biology·T FujimotoA Ukeshima
Jul 3, 1995·Proceedings of the National Academy of Sciences of the United States of America·W C SkarnesR S Beddington
Dec 10, 1999·Proceedings of the National Academy of Sciences of the United States of America·F Recillas-TargaG Felsenfeld
May 10, 2000·Proceedings of the National Academy of Sciences of the United States of America·A C GrothM P Calos
Feb 28, 2001·Reproduction : the Official Journal of the Society for the Study of Fertility·W M BaarendsJ A Grootegoed
May 22, 2001·Molecular and Cellular Biology·B ThyagarajanM P Calos
Aug 3, 2002·Proceedings of the National Academy of Sciences of the United States of America·Bonnie Burgess-BeusseGary Felsenfeld
Oct 16, 2002·Nature Biotechnology·Eric C OlivaresMichele P Calos
Jan 10, 2003·Nucleic Acids Research·D KarolchikUNKNOWN University of California Santa Cruz
Mar 26, 2003·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Judith L Leatherman, Thomas A Jongens
Apr 3, 2003·EMBO Reports·E Jane Albert Hubbard, Renee A Reijo Pera
Mar 27, 2004·Current Biology : CB·T Keith Blackwell
Dec 14, 2004·Nature·UNKNOWN International Chicken Genome Sequencing Consortium
Aug 30, 2005·Nature Biotechnology·Lei ZhuRobert J Etches
Sep 1, 2005·Proceedings of the National Academy of Sciences of the United States of America·Roland H FriedelMarc Tessier-Lavigne
Nov 22, 2005·Nature Methods·Bryan G Allen, Daniel L Weeks
Dec 6, 2005·Mechanisms of Development·Marie-Cecile van de LavoirRobert J Etches
May 9, 2006·Proceedings of the National Academy of Sciences of the United States of America·Mito Kanatsu-ShinoharaTakashi Shinohara
Jun 9, 2006·Nature·Marie-Cecile van de LavoirRobert J Etches
Nov 30, 2006·Cell·Geraldine Seydoux, Robert E Braun
Apr 10, 2007·Cell·Kathryn A O'Donnell, Jef D Boeke

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Citations

Jun 9, 2006·Nature·Marie-Cecile van de LavoirRobert J Etches
May 31, 2012·Proceedings of the National Academy of Sciences of the United States of America·Tae Sub Park, Jae Yong Han
Nov 28, 2013·Proceedings of the National Academy of Sciences of the United States of America·Benjamin SchusserPhilip A Leighton
May 26, 2012·PloS One·Marie-Cecile van de LavoirRobert J Etches
Jun 23, 2010·Yi chuan = Hereditas·Zhen-Hong SunRui-Liang Zhu
Apr 2, 2015·Poultry Science·Ellen CollariniRobert J Etches
May 14, 2016·Animal Science Journal = Nihon Chikusan Gakkaihō·Hiroshi Kagami
May 24, 2011·Preventive Veterinary Medicine·Wilfried A Kues, Heiner Niemann
May 16, 2012·Proceedings of the National Academy of Sciences of the United States of America·Joni MacdonaldMichael J McGrew
Feb 9, 2017·Development·Lorna TaylorMichael J McGrew
Nov 22, 2018·Poultry Science·Yi-Chen ChenShinn-Chih Wu
Jul 17, 2009·Biotechnology Journal·Götz Laible, Lucía Alonso-González
Jan 29, 2016·Transgenic Research·Timothy J DoranMark L V Tizard
Oct 3, 2019·Proceedings of the National Academy of Sciences of the United States of America·Mark E WoodcockMichael J McGrew
Mar 3, 2009·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Christopher T GordonPeter G Farlie
Jan 3, 2013·Development, Growth & Differentiation·Ken-ichi Nishijima, Shinji Iijima
Oct 26, 2018·Frontiers in Genetics·Hicham Sid, Benjamin Schusser
Jul 27, 2012·Cellular and Molecular Life Sciences : CMLS·Xiangyang Miao
Dec 18, 2013·Biology of Reproduction·Yonghong SongMarie-Cecile van de Lavoir
Jun 30, 2018·F1000Research·Ekaterina AntonovaPavel Volchkov
Oct 12, 2012·Annals of the New York Academy of Sciences·Tae Sub Park, Jae Yong Han
Mar 5, 2021·Proceedings of the National Academy of Sciences of the United States of America·Beate RieblingerBenjamin Schusser
Jun 24, 2010·Theriogenology·Sang Hyun ParkJae Yong Han
Oct 21, 2021·Biology of Reproduction·Nima DehdilaniHesam Dehghani

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