建立CRISPR-Cpf1辅助的基因编辑工具,以及在Corynebacterium glutamicum中对4-基素生物合成的工程
Rui Chen1,2, Feng Shi3,4,5, Youhe Xiang1,2
1State Key Laboratory of Food Science and Resources, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China.
World journal of microbiology & biotechnology
|July 31, 2023
概括
一个新的CRISPR-Cpf1系统提高了Corynebacterium glutamicum的基因编辑效率,将查率从5%提高到95%以上. 这种工具可以在代谢工程应用中促进4-基素 (4-HIL) 的产生.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 代谢工程是代谢工程.
背景情况:
- 科里尼细菌 (Corynebacterium glutamicum) 是一个重要的工业微生物,但由于基因组编辑方法效率低下而受到影响.
- 在C. glutamicum中现有的基因编辑技术具有局限性,阻碍了有效的基因修饰.
研究的目的:
- 开发一种改进的基因编辑系统,用于C. glutamicum.
- 在基因组编辑中提高第二交叉菌株的选效率.
- 应用新的代谢工程系统来生产4-氧化硫氨酸 (4-HIL).
主要方法:
- 设计并建立了一个包含CRISPR-Cpf1系统的pCS等离子体,该系统针对SacB系统的癌症基因.
- 利用CRISPR-Cpf1系统消除假阳性单个交叉菌株.
- 在4-HIL生产中应用了pCS辅助系统,用于促进器工程和前体供应优化.
主要成果:
- 这种pCS等离子体对携带kan基因的C. glutamicum菌株具有很高的杀伤性 (98.6%).
- 在PodhA::PilvBNC替代实验中,第二次交叉细菌的查效率从5%提高到95%以上.
- 工程菌株显示出增强的4-HIL产量,其中一个菌株达到137.0±33.9毫米.
- lysE 的减弱降低了 Lys 含量和 4-HIL 标位,证明了代谢途径的控制.
结论:
- 新的CRISPR-Cpf1辅助基因编辑工具显著提高了C. glutamicum的基因组编辑效率.
- 这种系统对代谢工程非常有效,能够增强像4-HIL.这样有价值的化合物的产生.
- 开发的工具为推进C. glutamicum的基因改造和代谢工程提供了有价值的解决方案.
更多相关视频
11:51Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
Published on: April 27, 2018
12.0K
07:46CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
Published on: December 11, 2020
5.9K
相关概念视频
01:28CRISPR/Cas9 Genome Editing
61
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
61
01:59CRISPR
52.4K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
52.4K