Development of specific TALENs for precision engineering in wheat

开发用于小麦精密工程的特定 TALEN

• 批准号: BB/L017687/1
• 负责人: Keith Edwards
• 金额: $ 10.5万
• 依托单位: University of Bristol
• 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%2FL017687%2F1
• 关键词: Development; specific; TALENs; precision; engineering

Recent developments in genome engineering mean that researchers now have tools which can be used to modify specific sequences within any species. One of these tools, called TALENs (transcription activator-like effector nucleases) could be of significant benefit to wheat researchers as they might allow plant geneticist and plant breeders to rapidly modify a crop to have unique agronomic properties while at the same time eliminating the need for non-host DNA to remain within the crop. Such improvements might include increased disease resistance, increased yield or better bread making qualities. While TALENs could revolutionise wheat breeding the system is not yet been fully tested, this is important as wheat is a polyploid (consisting of three different sets of chromosomes) and it has a large genome (consisting of approximately 5 times more DNA that the human genome). Hence, before TALENs can be applied to wheat, or any other crop, on a large scale, they need to be tested and monitored to confirm their early promise. Here we propose to test and monitor TALENs in wheat. To carry out this testing and monitoring we propose to utilise TALENs specific of two wheat genes, furthermore we propose to test the efficiency of TALEN activity when they are placed under the control of two promoters, one that is permanently switched on and one that can be induced by an increase in temperature.Following the introduction of the TALEN constructs into the wheat plant we will monitor their activity in terms of both their effectiveness at mutating the target genes and for NOT modifying other regions of the genome. Hence, one outcome of the project would be guidelines and protocols designed to ensure that researchers use the most effective promoters and gene sequences to increase ON target activity whilst simultaneously reducing or eliminating OFF target activity. Following successful completion of this preliminary study we will be well placed to advise government agencies on the effectiveness of TALENs and the steps that need to be put into place to ensure their safe use in future plant breeding.

基因组工程的最新发展意味着研究人员现在有了可以用来修改任何物种内特定序列的工具。其中一种被称为TALENs（转录激活因子样效应核酸酶）的工具可能对小麦研究人员有很大的好处，因为它们可能允许植物遗传学家和植物育种家快速修改作物，使其具有独特的农艺特性，同时消除了非宿主DNA留在作物内的需要。这些改进可能包括增强抗病性、提高产量或改善面包制作质量。虽然TALENs可能会彻底改变小麦育种，但该系统尚未经过全面测试，这很重要，因为小麦是多倍体（由三组不同的染色体组成），而且它有一个大的基因组（由大约5倍于人类基因组的DNA组成）。因此，在TALENs可以大规模应用于小麦或任何其他作物之前，需要对它们进行测试和监测，以确认它们的早期前景。在此，我们提出在小麦中进行TALENs的测试和监测。为了进行这种测试和监测，我们建议利用两个小麦基因的TALENs特异性，此外，我们建议测试两个启动子控制下TALEN活性的效率，一个启动子是永久开启的，另一个启动子可以通过温度升高诱导。在将TALEN构建体引入小麦植株后，我们将监测它们在突变靶基因和不修饰基因组其他区域方面的活性。因此，该项目的一个成果将是制定指导方针和方案，以确保研究人员使用最有效的启动子和基因序列来增加ON靶活性，同时减少或消除OFF靶活性。在这项初步研究成功完成后，我们将有能力就TALENs的有效性以及确保其在未来植物育种中安全使用所需采取的步骤向政府机构提供建议。

项目成果

CerealsDB 3.0: expansion of resources and data integration.

CEATEALSDB 3.0：资源和数据集成的扩展。

• DOI: 10.1186/s12859-016-1139-x
• 发表时间: 2016-06-24
• 期刊: BMC bioinformatics
• 影响因子: 3
• 作者: Wilkinson PA;Winfield MO;Barker GL;Tyrrell S;Bian X;Allen AM;Burridge A;Coghill JA;Waterfall C;Caccamo M;Davey RP;Edwards KJ
• 通讯作者: Edwards KJ

Wheat with greatly reduced accumulation of free asparagine in the grain, produced by CRISPR/Cas9 editing of asparagine synthetase gene TaASN2.

• DOI: 10.1111/pbi.13573
• 发表时间: 2021-08
• 期刊: Plant biotechnology journal
• 影响因子: 13.8
• 作者: Raffan S;Sparks C;Huttly A;Hyde L;Martignago D;Mead A;Hanley SJ;Wilkinson PA;Barker G;Edwards KJ;Curtis TY;Usher S;Kosik O;Halford NG
• 通讯作者: Halford NG