改进的原始编辑允许在Physcomitrium patens中进行常规可预测的基因编辑
Pierre-François Perroud1, Anouchka Guyon-Debast1, Josep M Casacuberta2
1Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin (IJPB), 78000 Versailles, France.
Journal of experimental botany
|May 27, 2023
概括
主编辑,精确的基因编辑工具,已被增强,以提高模型植物Physcomitrium patens.的效率. 这种优化的方法可以提高基因研究的编辑率,而不会影响准确度.
科学领域:
- 分子生物学分子生物学
- 植物科学 植物科学
- 遗传学 遗传学 是一个
背景情况:
- 有效和精确的基因编辑对于逆转基因学至关重要.
- 主编辑,CRISPR/Cas9衍生品,提供精度,但需要提高效率.
研究的目的:
- 改进主要编辑方法,以便在Physcomitrium patens.中常规使用.
- 调查提高主要编辑效率的策略,并探索其应用.
主要方法:
- 在Physcomitrium patens中标准化的原始质细胞转移.
- 评估各种主要编辑指导RNA (pegRNA) 结构和主要编辑器变体.
- 针对APT记者基因进行直接选择和Ppdek10进行间接选择.
主要成果:
- 优化主要编辑器表达和pegRNA修改显著增加了编辑率.
- 反转录酶模板中的同名突变提高了效率,但没有影响编辑质量.
- 通过间接选择证明了成功的基因编辑,并确定了用于主要编辑的植物逆转录酶.
- 展示了使用两个独立编码的的原始编辑潜力.
结论:
- 开发的方法允许在Physcomitrium patens.中进行常规和高效的prime编辑.
- 在主要编辑器组件和pegRNA设计的改进是改善编辑速率的关键.
- 主编辑是一种适用于植物直接和间接基因选择策略的多功能工具.
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