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没有DNA集成的CRISPR编辑的树植物的生成,使用Agrobacterium介导的转化技术

Shilin Sun¹, Xue Han¹, Ruoxuan Jin¹

¹College of Forestry, Shenyang Agricultural University, Shenyang 110866, China; Key Laboratory of Forest Tree Genetics, Breeding and Cultivation of Liaoning Province, Shenyang 110866, China.

Plant science : an international journal of experimental plant biology

|February 14, 2024

概括

这项研究引入了一种新的CRISPR编辑的DNA无植物生成方法,该方法使用树中短暂的Agrobacterium转化. 这种技术可以有效地生产基因组编辑的植物,而无需外来DNA集成,从而推进了植物生物技术.

科学领域:

植物生物技术植物生物技术
基因组编辑基因组编辑
分子生物学分子生物学

背景情况:

CRISPR/Cas9系统是一个强大的基因组编辑工具,但生成无DNA编辑植物仍然是一个重大挑战.
现有的方法经常与外来DNA集成扎,使下游应用程序和监管批准复杂化.

研究的目的:

开发和验证一种用于产生CRISPR编辑植物而无外来DNA集成的新方法,特别是Betula platyphylla (白).
提高植物基因组编辑技术的效率和适用性.

主要方法:

使用Agrobacterium介导的短暂遗传转换开发了CRISPR编辑的植物,没有外来DNA整合 (CPDAT) 方法.
引入T-DNA编码导向RNA (gRNA) 和Cas9进入树细胞以进行向DNA裂变.
从没有抗生素选择的转化扩展物诱导的偶然芽,允许在每个芽中独立检测突变.

主要成果:

在随机选择的线条中实现了80.00%的突变率,其中40.00%呈现双基突变.
成功识别了5个CRISPR编辑的子植物系 (7.69%) 没有外来DNA整合.
证明突变是保留和积累的,无论T-DNA的整合状态.

结论:

CPDAT方法提供了一种创新的策略,用于生产无DNA的CRISPR编辑的树植物,大大提高了效率.
这种方法为推进植物基因组编辑提供了有价值的工具,对各种植物物种具有广泛的潜力.
这些发现突出了开发更高效和多功能植物基因组编辑技术的有希望的方向.

关键词:

这种细菌名为Agrobacterium tumefaciens.树树树树克里斯普尔是什么意思?克里斯普尔是什么意思?没有DNA的植物是没有DNA的植物.

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