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Related Concept Videos

CRISPR and crRNAs02:53

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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.
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...
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Gene Regulation in Microbial Communities: Quorum Sensing01:28

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Quorum sensing is a mechanism of bacterial communication that enables coordinated gene expression in response to changes in population density. This facilitates collective behaviors that enhance survival, resource acquisition, and ecological adaptation. This process relies on small signaling molecules called autoinducers that accumulate as bacterial populations grow. When a critical threshold concentration of autoinducers is reached, bacterial cells collectively modify gene expression,...
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CRISPR/Cas9 Genome Editing01:28

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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...
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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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CRISPR01:59

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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...
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Related Experiment Video

Updated: Jul 13, 2025

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira
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Investigating Pseudomonas aeruginosa Gene Function During Pathogenesis Using Mobile-CRISPRi.

Michelle A Yu1, Amy B Banta2,3, Ryan D Ward2,4

  • 1Division of Pulmonary, Critical Care, Allergy, and Sleep Medicine, Department of Medicine, University of California, San Francisco, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 11, 2023
PubMed
Summary

We developed Mobile-CRISPRi, a tool for gene silencing in bacteria like P. aeruginosa. This system allows for partial gene knockdown without inducers, aiding research into essential genes and infection models.

Keywords:
CRISPR-Cas9CRISPRiEssential genesMurine pneumonia modelPseudomonas putidaPseudomonas syringaeSystems biology

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacterial Pathogenesis

Background:

  • CRISPR interference (CRISPRi) is a powerful gene silencing method.
  • Essential and conditionally essential genes in pathogens like P. aeruginosa influence antibiotic susceptibility and virulence.
  • Investigating these genes is crucial for understanding and combating bacterial infections.

Purpose of the Study:

  • To develop and optimize a versatile CRISPR interference (CRISPRi) system for gene silencing in diverse bacteria, including P. aeruginosa.
  • To create a tool, Mobile-CRISPRi, that facilitates the study of essential and conditionally essential genes.
  • To enable in vivo studies of gene function in P. aeruginosa infection models.

Main Methods:

  • Development of modular, mobilizable, and integrating vectors named Mobile-CRISPRi.
  • Optimization of Mobile-CRISPRi for constitutive low-level expression in P. aeruginosa, enabling inducible-free partial gene knockdown.
  • Establishment of protocols for creating Mobile-CRISPRi knockdown strains and evaluating phenotypes in a mouse pneumonia model.

Main Results:

  • Successful creation and validation of the Mobile-CRISPRi system for gene silencing in P. aeruginosa.
  • Demonstration of partial knockdown of essential and conditionally essential genes without exogenous inducers.
  • Application of the system in a P. aeruginosa mouse model of acute lung infection.

Conclusions:

  • Mobile-CRISPRi is an effective and adaptable tool for gene silencing in P. aeruginosa and other Pseudomonas species.
  • The system facilitates the investigation of essential and conditionally essential genes, contributing to the understanding of bacterial virulence and antibiotic resistance.
  • This technology supports in vivo studies of bacterial pathogenesis and the development of novel therapeutic strategies.