Efficient TALEN-mediated gene targeting of chicken primordial germ cells

Lorna Taylor; Daniel F. Carlson; Sunil Nandi; Adrian Sherman; Scott C. Fahrenkrug; Mike McGrew

doi:10.1242/dev.145367

Efficient TALEN-mediated gene targeting of chicken primordial germ cells

Lorna Taylor, Daniel F. Carlson, Sunil Nandi, Adrian Sherman, Scott C. Fahrenkrug, Mike McGrew

Royal (Dick) School of Veterinary Studies

Research output: Contribution to journal › Article › peer-review

Abstract

In this work we use TALE nucleases (TALENs) to target a reporter construct to the DDX4 (vasa) locus in chicken primordial germ cells. Vasa is a key germ cell determinant in many animal species and is posited to control avian germ cell formation. We show that TALENs mediate homology directed repair of the DDX4 locus on the Z sex chromosome at high (8.1%) efficiencies. Large genetic deletions of 30kb encompassing the entire DDX4 locus were also created using a single TALEN pair. The targeted PGCs were germ line competent and were used to produce DDX4 null offspring. In DDX4 knockout chickens, PGCs are initially formed but are lost during meiosis in the developing ovary leading to adult female sterility. TALEN-mediated gene targeting in avian primordial germ cells is therefore an efficient process.

Original language	English
Pages (from-to)	928-934
Journal	Development
Volume	144
Issue number	5
Early online date	7 Feb 2017
DOIs	https://doi.org/10.1242/dev.145367
Publication status	Published - 1 Mar 2017

Keywords / Materials (for Non-textual outputs)

TALEN
primordial germ cell
avian
transgenic knockout
chicken
ddx4

Access to Document

10.1242/dev.145367

Efficient TALEN-mediated gene targeting of chicken primordial germ cells
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
Final published version, 2.7 MBLicence: Creative Commons: Attribution (CC-BY)

National avian research facility
Sang, H. (Principal Investigator), Bernard, K. (Co-investigator), Hart, A. (Co-investigator), Kaiser, P. (Co-investigator), King, T. (Co-investigator) & Sherman, A. (Co-investigator)
Biotechnology and Biological Sciences Research Council
1/04/12 → 31/03/17
Project: Research
Control of development and reproductive traits
Burdon, T. (Principal Investigator), Argyle, D. (Co-investigator), Ashworth, C. (Co-investigator), Beard, P. (Co-investigator), Brunton, P. (Co-investigator), Burt, D. (Co-investigator), Clinton, M. (Co-investigator), Dunn, I. (Co-investigator), Farquharson, C. (Co-investigator), Headon, D. (Co-investigator), Hocking, P. (Co-investigator), Hohenstein, P. (Co-investigator), Hume, D. (Co-investigator), Jackson, I. (Co-investigator), McColl, B. (Co-investigator), McGrew, M. (Co-investigator), McLachlan, G. (Co-investigator), Sang, H. (Co-investigator), Summers, K. (Co-investigator) & Whitelaw, B. (Co-investigator)
Biotechnology and Biological Sciences Research Council
1/04/12 → 31/03/17
Project: Research

Mike McGrew
- mike.mcgrewroslin.ed.acuk
- Royal (Dick) School of Veterinary Studies - Personal Chair of Avian Reproductive Technologies
Person: Academic: Research Active

Cite this

@article{a867d79151b6415cabee3cf846994860,

title = "Efficient TALEN-mediated gene targeting of chicken primordial germ cells",

abstract = "In this work we use TALE nucleases (TALENs) to target a reporter construct to the DDX4 (vasa) locus in chicken primordial germ cells. Vasa is a key germ cell determinant in many animal species and is posited to control avian germ cell formation. We show that TALENs mediate homology directed repair of the DDX4 locus on the Z sex chromosome at high (8.1\%) efficiencies. Large genetic deletions of 30kb encompassing the entire DDX4 locus were also created using a single TALEN pair. The targeted PGCs were germ line competent and were used to produce DDX4 null offspring. In DDX4 knockout chickens, PGCs are initially formed but are lost during meiosis in the developing ovary leading to adult female sterility. TALEN-mediated gene targeting in avian primordial germ cells is therefore an efficient process. ",

keywords = "TALEN, primordial germ cell, avian, transgenic knockout, chicken, ddx4",

author = "Lorna Taylor and Carlson, \{Daniel F.\} and Sunil Nandi and Adrian Sherman and Fahrenkrug, \{Scott C.\} and Mike McGrew",

year = "2017",

month = mar,

day = "1",

doi = "10.1242/dev.145367",

language = "English",

volume = "144",

pages = "928--934",

journal = "Development",

issn = "0950-1991",

publisher = "Company of Biologists Ltd",

number = "5",

}

TY - JOUR

T1 - Efficient TALEN-mediated gene targeting of chicken primordial germ cells

AU - Taylor, Lorna

AU - Carlson, Daniel F.

AU - Nandi, Sunil

AU - Sherman, Adrian

AU - Fahrenkrug, Scott C.

AU - McGrew, Mike

PY - 2017/3/1

Y1 - 2017/3/1

N2 - In this work we use TALE nucleases (TALENs) to target a reporter construct to the DDX4 (vasa) locus in chicken primordial germ cells. Vasa is a key germ cell determinant in many animal species and is posited to control avian germ cell formation. We show that TALENs mediate homology directed repair of the DDX4 locus on the Z sex chromosome at high (8.1%) efficiencies. Large genetic deletions of 30kb encompassing the entire DDX4 locus were also created using a single TALEN pair. The targeted PGCs were germ line competent and were used to produce DDX4 null offspring. In DDX4 knockout chickens, PGCs are initially formed but are lost during meiosis in the developing ovary leading to adult female sterility. TALEN-mediated gene targeting in avian primordial germ cells is therefore an efficient process.

AB - In this work we use TALE nucleases (TALENs) to target a reporter construct to the DDX4 (vasa) locus in chicken primordial germ cells. Vasa is a key germ cell determinant in many animal species and is posited to control avian germ cell formation. We show that TALENs mediate homology directed repair of the DDX4 locus on the Z sex chromosome at high (8.1%) efficiencies. Large genetic deletions of 30kb encompassing the entire DDX4 locus were also created using a single TALEN pair. The targeted PGCs were germ line competent and were used to produce DDX4 null offspring. In DDX4 knockout chickens, PGCs are initially formed but are lost during meiosis in the developing ovary leading to adult female sterility. TALEN-mediated gene targeting in avian primordial germ cells is therefore an efficient process.

KW - TALEN

KW - primordial germ cell

KW - avian

KW - transgenic knockout

KW - chicken

KW - ddx4

U2 - 10.1242/dev.145367

DO - 10.1242/dev.145367

M3 - Article

SN - 0950-1991

VL - 144

SP - 928

EP - 934

JO - Development

JF - Development

IS - 5

ER -

Efficient TALEN-mediated gene targeting of chicken primordial germ cells

Abstract

Keywords / Materials (for Non-textual outputs)

Access to Document

Fingerprint

Projects

National avian research facility

Control of development and reproductive traits

Profiles

Mike McGrew

Cite this