建立CRISPRi用于可编程基因抑制和基因组进化在Cupriavidus necator中
Zhijiao Wang1,2, Haojie Pan1,2, Sulin Ni1
1Key Laboratory of Biomass Chemical Engineering of Ministry of Education & National Key Laboratory of Biobased Transportation Fuel Technology, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
ACS synthetic biology
|February 13, 2024
概括
我们开发了一种优化的CRISPR干扰 (CRISPRi) 系统,用于Cupriavidus necator中的基因抑制. 该工具通过实现精确的基因控制和快速菌株进化来增强生物技术的微生物细胞工厂.
科学领域:
- 微生物学 微生物学
- 合成生物学 合成生物学
- 生物技术是生物技术.
背景情况:
- 库普里亚维杜斯 (Cupriavidus necator) H16是一种多功能微生物细胞工厂,广泛利用碳来源.
- 有限的基因组工程工具,特别是基因调节,阻碍了它的全部潜力.
- 克里斯普尔/Cas9技术为先进的基因操纵提供了一个有前途的解决方案.
研究的目的:
- 开发和优化CRISPR干扰 (CRISPRi) 系统,用于C. necator. 的基因抑制.
- 为了证明CRISPRi对代谢工程和基因组进化的有用性.
- 为C. necator细胞工厂优化提供一个高效的工具包.
主要方法:
- 在C. necator.中表达一种编码子优化的失活的Cas9 (dCas9) 和单向导RNA (sgRNAs).
- 应用CRISPRi用于抑制外源和内源基因.
- 案例研究涉及聚基酸盐 (PHB) 减少,烯合成和CRISPRi-Mutator系统的建立.
主要成果:
- 一个优化的CRISPRi系统在C. necator.中成功建立和验证.
- 克里斯皮尔使可编程和可控制的基因抑制成为可能.
- 代谢流量从PHB重新连接到利科合成.
- 克里斯皮尔i-突变器促进了微生物电合成 (MES) 强壮突变菌株的快速生成.
结论:
- 开发的CRISPRi系统是用于C. necator.基因操纵的高效和多功能工具.
- 这套工具提升了C. necator作为微生物生产平台的优化.
- 这些发现通过增强细胞工厂能力来支持各种生物技术应用.
关键词:
克里斯普里是什么?克里斯普里是什么?鱼虫 (Cupriavidus necator) H16 H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) H16 鱼虫 (Cupriavidus necator) 鱼虫 (Cupria基因组的演化 基因组的演化利科生物合成的生物合成代谢工程是代谢工程.相关概念视频
CRISPR and crRNAs
17.0K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
17.0K
Cis-regulatory Sequences
9.9K
Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...
9.9K
CRISPR
51.0K
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...
51.0K
Conservative 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


