Gene targeting of primordial germ cells using TALEN technology to generate the first knockout transgenic chickens

利用TALEN技术对原始生殖细胞进行基因打靶，培育出第一批基因敲除转基因鸡

• 批准号: BB/L018063/1
• 负责人: Michael McGrew
• 金额: $ 13.03万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL018063%2F1
• 关键词: Gene; targeting; primordial; germ; cells

The chicken is used to study how embryos form and is also a major agricultural species. Being able to selectively target genes in chickens will help to understand and improve disease resistance in birds and poultry production. This will also give us new ways to study how chicken embryos form. Sex stem cells can be used to precisely modify the chicken genome. Gene targeting using TALE nucleases (TALENs) promises to be an efficient method to modify sex stem cells to make knockout chickens. We propose to develop this novel technology to efficiently target genes in sex stem cells using TALEN-directed homologous recombination. In this proposal, we will target two genes needed for germ cells to form. Targeted sex stem cells will be injected into host embryos and bred to produce targeted knockout chickens. The knockout chicken produced will be useful to understand how germ cells form and also as a recipient for sex stem cells from frozen avian biobanks for domesticated poultry and potentially for endangered avian species. These advances will support research in basic biology of vertebrate development with applications in understanding disease resistance and fertility in poultry.

鸡被用来研究胚胎如何形成，也是一种主要的农业物种。能够选择性地靶向鸡的基因将有助于了解和提高鸟类和家禽生产的抗病能力。这也将为我们研究鸡胚胎如何形成提供新的方法。性干细胞可用于精确修改鸡的基因组。利用TALE核酸酶（TALENs）进行基因打靶有望成为修饰性干细胞以制造基因敲除鸡的有效方法。我们建议开发这种新技术，使用TALEN指导的同源重组有效地靶向性干细胞中的基因。在这项提案中，我们将针对生殖细胞形成所需的两个基因。有针对性的性干细胞将被注射到宿主胚胎中，并培育出有针对性的基因敲除鸡。生产的基因敲除鸡将有助于了解生殖细胞如何形成，也可以作为来自冷冻禽类生物库的性干细胞的受体，用于驯养家禽和潜在的濒危鸟类物种。这些进展将支持脊椎动物发育的基础生物学研究，并应用于了解家禽的抗病性和生育能力。

项目成果

High fidelity CRISPR/Cas9 increases precise monoallelic and biallelic editing events in primordial germ cells.

• DOI: 10.1038/s41598-018-33244-x
• 发表时间: 2018-10-11
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Idoko-Akoh A;Taylor L;Sang HM;McGrew MJ
• 通讯作者: McGrew MJ

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

• DOI: 10.1242/dev.145367
• 发表时间: 2017-03-01
• 期刊: Development (Cambridge, England)
• 作者: Taylor L;Carlson DF;Nandi S;Sherman A;Fahrenkrug SC;McGrew MJ
• 通讯作者: McGrew MJ

Gene editing in birds takes flight.

鸟类中的基因编辑可以飞行。

• DOI: 10.1007/s00335-017-9701-z
• 发表时间: 2017-08
• 期刊: Mammalian genome : official journal of the International Mammalian Genome Society
• 作者: Woodcock ME;Idoko-Akoh A;McGrew MJ
• 通讯作者: McGrew MJ

FGF, Insulin, and SMAD Signaling Cooperate for Avian Primordial Germ Cell Self-Renewal.

• DOI: 10.1016/j.stemcr.2015.10.008
• 发表时间: 2015-12-08
• 期刊: Stem cell reports
• 影响因子: 5.9
• 作者: Whyte J;Glover JD;Woodcock M;Brzeszczynska J;Taylor L;Sherman A;Kaiser P;McGrew MJ
• 通讯作者: McGrew MJ