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通过优化日本雪松 (Cryptomeria japonica D. Don) 的Cas9编码子来提高基因组编辑效率

Yoshihiko Nanasato¹, Harunori Kawabe¹, Saneyoshi Ueno²

¹Forest Bio-Research Center, Forestry and Forest Products Research Institute (FFPRI), Forest Research and Management Organization (FRMO), 3809-1 Ishi, Juo, Hitachi, Ibaraki 319-1301, Japan.

Plant biotechnology (Tokyo, Japan)

|March 14, 2025

概括

我们开发了一种编码子优化的Cas9基因 (CjSpCas9),以提高日本雪松 (Cryptomeria japonica) 的CRISPR/Cas9基因组编辑效率. 这种增强的系统显著提高了双基突变率,加速了针叶树的分子繁殖.

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
针叶研究针叶研究

背景情况:

日本香柏 (Cryptomeria japonica) 是日本的一个重要的种植针叶树.
现有的CRISPR/Cas9系统在C. japonica中对双基突变的效率很低.

研究的目的:

为了提高CRISPR/Cas9基因组编辑效率在日本香柏树中.
为了改善基因淘汰的双基突变的产生.

主要方法:

对C. japonica. 的Cas9基因的优化.
使用CjChl I基因对编辑效率的评估.
与优化用于Arabidopsis thaliana (AtSpCas9) 和Oryza sativa (OsSpCas9) 的Cas9变种进行比较.
西方斑点检查以确认Cas9蛋白水平.

主要成果:

与AtSpCas9和OsSpCas9.9相比,对CjChl I而言,编码子优化的CjSpCas9实现了显著更高的双干扰效率.
对目标t4来说,双干扰效率高出1.8-2.0倍,对目标t1+t2.4来说高出4.9-1.4倍.
通过西部涂抹证实了增加的Cas9蛋白积累,与更好的编辑效率相关.

结论:

优化Cas9的Codon是有效提高基因组编辑效率在日本香柏树.
开发的CjSpCas9系统加速了双基突变的产生.
这种改进的系统将促进针叶树的分子繁殖.

关键词:

这就是CRISPR/Cas9的作用.这种病名为Cryptomeria japonica.日本香柏树是日本的香柏树.代码子优化代码子优化基因组编辑基因组编辑

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