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Updated: May 27, 2025

Ubiquitous and Tissue-specific RNA Targeting in Drosophila Melanogaster using CRISPR/CasRx
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Reprogrammable RNA-targeting CRISPR systems evolved from RNA toxin-antitoxins.

Shai Zilberzwige-Tal1, Han Altae-Tran2, Soumya Kannan1

  • 1Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research at MIT, Cambridge, MA 02139, USA; Department of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

Cell
|February 19, 2025
PubMed
Summary
This summary is machine-generated.

Researchers traced the evolution of RNA-targeting CRISPR-Cas13 systems, finding they likely originated from abortive infection (AbiF) toxin-antitoxin systems. This study reveals key structural changes enabling this evolutionary transition.

Keywords:
CRISPRCas13RNA-guided mechanismbacterial immunitytoxin-antitoxin system

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Evolutionary Biology

Background:

  • CRISPR systems are crucial for adaptive immunity in prokaryotes.
  • The evolutionary pathways leading to RNA-guided CRISPR systems, particularly Cas13, are not fully understood.
  • Understanding CRISPR evolution provides insights into genome defense mechanisms.

Purpose of the Study:

  • To identify the evolutionary ancestors of the RNA-targeting CRISPR-Cas13 system.
  • To characterize the evolutionary intermediate Cas13e and its relationship with AbiF.
  • To elucidate the structural and functional divergence between AbiF and Cas13 systems.

Main Methods:

  • Integrated sequence and structure evolutionary tracing.
  • Biochemical characterization of Cas13e and AbiF.
  • Cryo-electron microscopy (cryo-EM) for structure determination of AbiF.

Main Results:

  • Cas13 systems likely evolved from AbiF, a toxin-antitoxin system linked to abortive infection.
  • A miniature Cas13, termed Cas13e, was identified as an evolutionary intermediate.
  • AbiF functions as a toxin-antitoxin system with an RNA antitoxin, distinct from Cas13's RNA-guided RNA targeting.
  • Cryo-EM structure of AbiF revealed structural differences compared to Cas13s.
  • Key structural alterations facilitating the evolution from AbiF to CRISPR were mapped.

Conclusions:

  • The study provides a clear evolutionary trajectory for RNA-targeting CRISPR-Cas13 systems.
  • AbiF represents a plausible precursor to Cas13, highlighting functional and structural transitions.
  • The findings deepen our understanding of the evolution of programmable RNA-guided systems.