Structural basis for target DNA cleavage and guide RNA processing by CRISPR-Casλ2

Jun 5, 2025Communications biology

How CRISPR-Casλ2 cuts target DNA and processes its guide RNA

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Abstract

The cryo-electron microscopy structures of Casλ2 revealed five distinct functional states.

  • Casλ2 demonstrated nuclease activity in both mammalian and plant cells, indicating its potential for genome editing.
  • The study identified a unique ruler mechanism by which Casλ2 processes precursor crRNA to mature crRNA.
  • Dynamic domain rearrangements during Casλ2 activation were observed, providing insights into its molecular mechanisms.
  • Structural comparisons with Casλ1 and CasΦ highlighted both diversity and conservation among phage-encoded type V CRISPR-Cas enzymes.
  • These findings enhance the understanding of CRISPR-Cas nucleases and may aid in the engineering of CRISPR-Casλ-based genome-editing tools.

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Key numbers

16–18 nt
Optimal Spacer Length for Cleavage
Casλ2 cleaves DNA optimally with guide RNAs of this length.
16–18 nt
Spacer Length of Mature crRNA
Casλ2 processes to mature crRNA of this length.

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