在Streptococcus agalactiae多部位序列17型菌株中的CRISPR干扰
bioRxiv : the preprint server for biology
|September 5, 2025
概括
研究人员为B组链球菌 (GBS) ST-17菌株开发了CRISPR干扰系统. 这种工具可以通过向基因敲除来研究GBS脑膜炎在血脑屏障上的致病性.
科学领域:
- 微生物学
- 遗传学
- 传染性疾病
背景情况:
- B型链球菌 (GBS) 是新生儿细菌性脑膜炎的主要原因.
- 像COH1一样,高病毒性血清型III,序列型17 (ST-17) 的GBS菌株与严重的新生儿疾病有很强的联系.
- 对ST-17GBS菌株进行基因操作是很困难的,这阻碍了对毒性因素的研究.
研究的目的:
- 在ST-17 GBS COH1菌株中开发CRISPR干扰 (CRISPRi) 系统.
- 为了使功能基因组学和高通量查GBS毒性因素.
- 促进血脑屏障的GBS病变研究.
主要方法:
- 在COH1 GBS菌株中使用催化无活化Cas9 (dCas9) 的CRISPR干扰系统的开发.
- 使用血液溶解试验,qPCR和使用人脑内皮细胞的体外感染模型来证实系统的有效性.
- 针对性打击关键病毒性基因,包括pilA,srr2和iagA.
主要成果:
- 在ST-17 GBS COH1中成功实现可调整的CRISPRi系统.
- 证明了基本的GBS毒性基因的表型破坏.
- 减少了血脑屏障中的细菌粘附,入侵和炎症反应.
结论:
- 开发的CRISPRi系统为ST-17 GBS的快速基因淘汰提供了一个多功能平台.
- 这种工具克服了COH1的先前基因操纵挑战.
- 能够对血脑屏障中的GBS病变和宿主病原体相互作用进行先进的研究.
相关概念视频
CRISPR and crRNAs
17.3K
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...
17.3K
The Antiviral System of Bacteria and Archaea: CRISPR
121
CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
121
CRISPR/Cas9 Genome Editing
208
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...
208
CRISPR
52.8K
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...
52.8K
Conservative Site-specific Recombination and Phase Variation
6.1K
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...
6.1K


