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在植物中使用可诱导热冲击的基因组编辑系统进行临时基因淘汰

Zhen Liang¹, Sha Wei¹, Yuqing Wu¹

¹School of Life Science, Shanxi University, Taiyuan, Shanxi, China.

The plant genome

|August 2, 2023

概括

研究人员开发了两种热冲击诱导基因组编辑 (IGE) 系统,用于精确的植物基因功能研究. HSP18.2 IGE系统在不同发育阶段的基因编辑中表现出卓越的效率.

科学领域:

植物分子生物学植物分子生物学
基因组编辑技术的技术.
功能性基因组学是一种功能性基因组学.

背景情况:

克里斯普尔/Cas9是产生功能丧失突变的关键工具.
需要对基因编辑进行精确的空间和时间控制,以进一步剖析基因功能.
条件基因组编辑系统对于研究基本基因和发育过程至关重要.

研究的目的:

开发和表征热冲击诱导基因组编辑 (IGE) 系统,用于在Arabidopsis thaliana中进行条件基因修饰.
为了比较天然 (HSP18.2) 和合成 (HSE-COR15A) 热诱导促进剂在驱动Cas9表达的效率.
评估IGE系统在特定发育阶段对基因失活的有用性.

主要方法:

使用可热诱导的促进子 (HSP18.2和HSE-COR15A) 构建了两个IGE系统,以驱动Cas9表达.
在Arabidopsis thaliana中测试了系统,在播种和螺栓化阶段使用循环和连续的热冲击处理.
通过准BRI1和PDS基因来评估基因编辑效率,并使用qRT-PCR量化Cas9表达.

主要成果:

在循环和连续热处理下,HSP18.2 IGE 系统显示出比 HSE-COR15A 系统更高的 Cas9 表达和更有效的突变发生.
这两种系统都成功地诱导了幼苗阶段的基因编辑,但只有HSP18.2在螺栓化阶段有效.
针对BRI1和PDS的向突变产生了明确的表型结果,证实了IGE系统的功能.

结论:

热冲击诱导基因组编辑系统为植物条件基因失活提供了一种强大的方法.
HSP18.2促进体提供了强大的和高效的热诱导Cas9表达,用于跨发育阶段的精确基因组编辑.
这些IGE系统是剖析基本基因功能和了解植物发育生物学的宝贵工具.

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