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Related Concept Videos

CRISPR and crRNAs02:53

CRISPR and crRNAs

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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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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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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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CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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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Defense Against Bacterial Pathogens01:31

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The human immune system is a complex network of cells, tissues, and organs that work together to defend the body against bacterial infections. It consists of various immune cells, each playing a specific role in the defense mechanism.
Phagocytes
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CRISPR01:59

CRISPR

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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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Gene Regulation in Microbial Communities: Quorum Sensing01:28

Gene Regulation in Microbial Communities: Quorum Sensing

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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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Related Experiment Video

Updated: Oct 25, 2025

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira
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CRISPR interference identifies vulnerable cellular pathways with bactericidal phenotypes in Mycobacterium

Matthew B McNeil1,2, Laura M Keighley1, Josephine R Cook1

  • 1Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand.

Molecular Microbiology
|August 4, 2021
PubMed
Summary

New research uses CRISPR interference to identify essential drug targets in Mycobacterium tuberculosis. This approach distinguishes gene vulnerability and lethality, aiding antibiotic development for this leading cause of death.

Keywords:
CRISPR interferencemycobacteriatuberculosis

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A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening
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Area of Science:

  • Microbiology
  • Genetics
  • Drug Discovery

Background:

  • Mycobacterium tuberculosis is a major global health threat necessitating novel drug development.
  • Current genetic strategies for identifying drug targets have limitations in assessing gene essentiality and lethality.
  • Distinguishing between essential, vulnerable, and lethal genes is crucial for effective antibiotic target prioritization.

Purpose of the Study:

  • To investigate the essentiality, vulnerability, and lethality of 94 target genes in Mycobacterium tuberculosis.
  • To overcome limitations of existing genetic deletion strategies using a novel CRISPR interference approach.
  • To identify and prioritize potential antibiotic targets by understanding their role in bacterial survival and death.

Main Methods:

  • Utilized mycobacterial CRISPR interference (CRISPRi) for targeted gene knockdown.
  • Combined CRISPRi with high-throughput phenotypic screening.
  • Assessed the essentiality, vulnerability, and lethality of 94 diverse target genes across various cellular pathways.

Main Results:

  • Essential genes in cell wall synthesis and central cellular functions were highly vulnerable, often leading to bactericidal effects.
  • Genes involved in metabolism, oxidative phosphorylation, and amino acid synthesis showed lower vulnerability and frequently resulted in bacteriostatic effects.
  • Successfully differentiated levels of gene essentiality and lethality, providing a scalable molecular tool.

Conclusions:

  • The study provides a refined method for assessing gene essentiality and lethality in Mycobacterium tuberculosis.
  • Findings offer novel insights into mycobacterial genetics and biology, crucial for prioritizing antibiotic drug targets.
  • This research aids in the development of new strategies to combat tuberculosis by identifying key vulnerabilities.