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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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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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A PNPase dependent CRISPR System in Listeria.

Nina Sesto1, Marie Touchon2, José Marques Andrade3

  • 1Unité des Interactions Bactéries-Cellules, Institut Pasteur, Paris, France ; INSERM, U604, Paris, France ; INRA, USC2020, Paris, France.

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Summary

This study reveals a novel CRISPR system in Listeria monocytogenes that functions without cas genes, relying instead on the PNPase enzyme for processing and DNA interference against phages.

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Area of Science:

  • * Microbiology
  • * Molecular Biology
  • * Genetics

Background:

  • * CRISPR (clustered regularly interspaced short palindromic repeats) systems provide bacterial defense against mobile genetic elements.
  • * CRISPR-associated (cas) genes are typically essential for CRISPR function.
  • * Listeria monocytogenes is a significant bacterial pathogen and a model for RNA regulation.

Purpose of the Study:

  • * To characterize a novel CRISPR element (RliB) in Listeria monocytogenes.
  • * To investigate the processing and function of this cas-less CRISPR system.
  • * To explore the role of polynucleotide phosphorylase (PNPase) in CRISPR activity.

Main Methods:

  • * Structural probing to determine RliB secondary structure.
  • * Genomic analysis to assess RliB distribution and association with cas genes.
  • * Plasmid acquisition assays to evaluate RliB interference.
  • * Genetic manipulation to assess the requirement of PNPase for RliB function.

Main Results:

  • * RliB exhibits an unusual secondary structure with repeat-spacer basepairing.
  • * RliB is conserved across Listeria genomes, independent of cas genes.
  • * RliB functions as a substrate for PNPase, which is crucial for its processing and DNA interference.
  • * Cas-less RliB reduces plasmid acquisition when trans-encoded cas genes are present.

Conclusions:

  • * A novel, cas-less CRISPR system (RliB) has been identified in Listeria.
  • * CRISPR processing and DNA interference are dependent on the host enzyme PNPase.
  • * This discovery expands the known mechanisms of CRISPR-mediated immunity and highlights a new role for PNPase in CRISPRology.