在Fusarium fujikuroi中建立用于代基因编辑的可选择标记回收系统
Tian-Qiong Shi1,2, Cai-Ling Yang2, Dong-Xun Li2
1Jiangxi New Reyphon Biochemical Co., Ltd, Salt & Chemical Industry, Xingan, Jiangxi, 331399, People's Republic of China.
Synthetic and systems biotechnology
|February 9, 2024
概括
研究人员为Fusarium fujikuroi开发了一种新的选择标记回收系统,使得有效的基因编辑成为可能. 这一进步促进了代谢工程,以改善农业中的吉伯酸 (GA3) 生产.
科学领域:
- 生物技术是生物技术.
- 合成生物学 合成生物学
- 植物科学 植物科学
背景情况:
- 吉伯酸 (GA3) 是农业中重要的植物生长激素,通过使用Fusarium fujikuroi的发酵产生.
- 目前F. fujikuroi的代谢工程受到缺乏有效的选择标记回收系统的限制,阻碍了代基因编辑.
研究的目的:
- 开发和验证F. fujikuroi的选择标记回收策略.
- 为了实现高效的代基因编辑,增强GA3的生产.
主要方法:
- 开发了一种与CRISPR/Cas9相结合的pyr4爆破器系统,用于回收5'-酸盐脱碳酶 (Pyr4) 选择标记物.
- 应用了F. fujikuroi的代基因整合和大基因删除系统.
- 通过过度表达关键酶:geranylgeranyl pyrophosphate synthase和截断的3-hydroxy-3-methyl glutaryl 共酶 A 减少酶来设计GA3 生物合成.
主要成果:
- 在F. fujikuroi中成功展示了用于标记物回收的pyr4-blaster系统.
- 实现了高效的代基因编辑,包括大基因删除.
- 通过新陈代谢途径重新连接,显著改善GA3标位.
结论:
- 已建立的选择标记回收系统对于F. fujikuroi的代基因编辑非常有效.
- 该系统为先进的代谢工程策略提供了基础,以优化工业GA3生产.
相关概念视频
02:53Conservative Site-specific Recombination and Phase Variation
6.0K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
6.0K
01:16In-vitro Mutagenesis
13.9K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
13.9K