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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.
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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...
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Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira
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CRISPR interference in a Streptococcus agalactiae multi-locus sequence type 17 strain.

William D Cutts1, Aidan W Flanagan2, Brice K Gorman2

  • 1Department of Molecular and Cell Biology, University of Texas at Dallas, Dallas, Texas, USA.

Journal of Bacteriology
|January 14, 2026
PubMed
Summary
This summary is machine-generated.

Researchers developed a CRISPR interference (CRISPRi) system to study Group B Streptococcus (GBS) pathogenesis. This tool allows for targeted gene knockdown in the hypervirulent ST-17 strain COH1, aiding research into neonatal meningitis.

Keywords:
CRISPRGroup B Streptococcus

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Area of Science:

  • Microbiology
  • Genetics
  • Infectious Diseases

Background:

  • Group B Streptococcus (GBS) is a primary cause of neonatal bacterial meningitis.
  • The hypervirulent serotype III, sequence type 17 (ST-17) strain COH1 is linked to severe disease but is genetically challenging to manipulate.
  • Understanding GBS virulence factors at the blood-brain barrier (BBB) is crucial for combating neonatal meningitis.

Purpose of the Study:

  • To develop a novel CRISPR interference (CRISPRi) system for targeted gene knockdown in the GBS ST-17 COH1 strain.
  • To enable functional genomics and high-throughput screening of GBS virulence factors.
  • To investigate GBS interactions at the BBB.

Main Methods:

  • Development of a CRISPR interference (CRISPRi) system using catalytically inactivated Cas9 (dCas9) in the COH1 strain.
  • Confirmation of system efficacy via hemolysis assays and qPCR transcriptional analysis.
  • Assessment of gene knockdown effects in vitro using human brain endothelial cell infection models.

Main Results:

  • The CRISPRi system successfully achieved tunable gene expression knockdown in ST-17 GBS.
  • Phenotypic knockdowns of key virulence genes (PI-2b, srr2, iagA) were observed.
  • Reduced bacterial adhesion, invasion, and inflammatory responses at the BBB were demonstrated.

Conclusions:

  • The developed CRISPRi platform provides a versatile tool for genetic manipulation in ST-17 GBS.
  • This system facilitates rapid functional genomics and pathogenesis research for GBS.
  • The findings contribute to a better understanding of GBS virulence and potential therapeutic targets.