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Updated: Feb 13, 2026

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Scalable Transfection of Maize Mesophyll Protoplasts
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从玉米中细胞中分离和转染原生质
Lauren A Higa1, Taren Bouwman1, Zhi-Yan Du1
1Department of Molecular Biosciences & Bioengineering, University of Hawaii at Mānoa, Honolulu, HI, USA.
Bio-protocol
|February 12, 2026
概括
这项研究提出了玉米原质细胞分离和CRISPR/Cas9基因编辑的优化协议. 这种新方法提高了原始塑料的产量,活力和转化效率,加速了植物研究.
科学领域:
- 植物分子生物学 植物分子生物学
- 农业生物技术 农业生物技术
背景情况:
- 原生体系统对于植物研究和基因编辑验证至关重要.
- 通过不成熟的胚胎转化玉米是缓慢的,资源密集的.
- 现有的玉米原型塑料协议在恢复,转染和可活性方面缺乏一致性.
研究的目的:
- 开发一个优化的协议,用于玉米 mesophyll 原始细胞的隔离和PEG介导的转染.
- 提高玉米基因编辑评估的效率,可复制性和速度.
主要方法:
- 优化了使用垂直切割从etiolated幼苗中对玉米 mesophyll 原塑体的隔离.
- 使用CRISPR/Cas9构造的PEG4000介导的转染,使用减少的等离子体DNA (10μg).
- 修改了洗和储存条件,以延长原生质细胞的活力长达七天.
主要成果:
- 通过垂直切割提高了原生质的产量,一致性和活力.
- 有效的CRISPR/Cas9传递仅使用10μg的等离子体DNA.
- 将转化原生质的生存时间延长到七天,以进行增强的下游分析.
- 使用目标测序和Cas-Analyzer对编辑效率的量化.
结论:
- 该协议提供了一种快速,高效和可重复的方法来评估玉米中的基因编辑.
- 这种方法加速了农作物研究和植物分子生物学研究.
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