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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.
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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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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: Oct 27, 2025

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira
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A Cas12a-based CRISPR interference system for multigene regulation in mycobacteria.

Neil Fleck1, Christoph Grundner2

  • 1Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, Washington, USA.

The Journal of Biological Chemistry
|July 23, 2021
PubMed
Summary

Researchers developed a new Cas12a-based CRISPR interference (CRISPRi) system for mycobacteria. This advanced tool enables efficient, simultaneous gene regulation in bacteria, even within infected cells, aiding the study of essential genes.

Keywords:
CRISPRiCas12aMycobacterium tuberculosisgene regulationmycobacteria

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Mycobacteria pose a significant global health challenge.
  • Genetic manipulation of mycobacteria has historically been difficult.
  • Existing CRISPR interference (CRISPRi) systems, like Cas9-based platforms, have limitations in multigene regulation.

Purpose of the Study:

  • To introduce a novel CRISPRi platform for mycobacteria utilizing the Cas12a enzyme.
  • To overcome limitations of previous CRISPRi systems for efficient gene repression.
  • To provide a versatile tool for studying mycobacterial genetics and enabling new therapeutic strategies.

Main Methods:

  • Development of a synthetic CRISPR array system with the minimal type V Cas12a enzyme.
  • Application of the Cas12a-based CRISPRi system for gene regulation in vitro.
  • Testing the system's efficacy in regulating essential and multiple genes simultaneously.
  • Evaluation of the system's performance in infected macrophages.

Main Results:

  • The Cas12a-based CRISPRi system demonstrates high efficiency in gene repression in mycobacteria.
  • The system allows for tunable and reversible gene regulation.
  • Simultaneous regulation of multiple genes, including essential ones, is achieved.
  • The platform functions effectively both in vitro and within infected host cells.

Conclusions:

  • Cas12a-based CRISPRi offers a simplified and powerful method for genetic manipulation in mycobacteria.
  • This tool facilitates the investigation of complex genetic interactions and conditional gene essentiality during infection.
  • The system holds promise for identifying novel drug targets, particularly synthetically lethal ones, against Mycobacterium tuberculosis (Mtb).