超声波辅助的胡须方法使CRISPR-Cas9核糖蛋白传递能够诱导大米中的基因组编辑
Akiyoshi Nakamura1, Tsubasa Yano2, Nobutaka Mitsuda1,3
1Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1, Higashi, Tsukuba, Japan.
Scientific reports
|September 7, 2023
概括
研究人员开发了一种新的CRISPR/Cas9核糖蛋白 (RNP) 输送方法,用于使用超声波辅助. 这种技术可以在植物中进行高效的基因组编辑,避免工业应用的转基因整合.
科学领域:
- 植物生物技术 植物生物技术
- 分子生物学分子生物学
- 农业科学 农业科学
背景情况:
- 克里斯普尔/卡斯9技术为植物育种提供了巨大的潜力.
- 将基因组编辑工具输入植物细胞是具有挑战性的,因为它们的细胞壁很硬.
- 避免转基因插入对于工业工厂应用至关重要,这使得核蛋白 (RNP) 输送是可取的.
研究的目的:
- 介绍一种用于将Cas9-gRNARNARNAP输入米细胞的新方法.
- 建立一个高效的RNP交付系统,用于生成基因组编辑植物.
- 为了证明超声波辅助胡须方法在大米中RNP传递的实用性.
主要方法:
- 适应超声波辅助胡须方法,用于在大米中输送RNP.
- 与选择的标记基因一起提供RNP.
- 从经过处理的木头中再生编辑的植物组织.
主要成果:
- 通过RNP交付生成的OsLCYβ基因组编辑大米系的成功隔离.
- 观察到在OsLCYβ突变的calli和芽中异常的胡卜素积累.
- 通过使用超声波辅助胡须方法通过RNP传递检测到米中的基因组编辑事件,即使在低频率.
结论:
- 超声波辅助胡须方法是一种可行的方法,可以将RNP输送到米细胞中.
- 这种方法有助于创造无转基因,基因组编辑的植物.
- 开发的技术有望推进使用CRISPR/Cas9技术的植物育种应用.
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