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Updated: Aug 23, 2025
09:51
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
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Broad-spectrum CRISPR-Cas13a enables efficient phage genome editing.
Benjamin A Adler1,2, Tomas Hessler2,3,4, Brady F Cress1,2
1California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, CA, USA.
Nature Microbiology
|November 1, 2022
Summary
Leptotrichia buccalis Cas13a (LbuCas13a) demonstrates potent antiviral activity against diverse phages. This CRISPR-Cas13 system enables efficient, broad-spectrum phage genome editing, including multi-gene deletions.
Area of Science:
- Molecular Biology
- Microbiology
- Genetics
Background:
- CRISPR-Cas13 systems are RNA-guided RNA nucleases with natural antiviral defense mechanisms.
- These systems target foreign RNA, leading to its degradation and inhibiting pathogen replication.
Purpose of the Study:
- To analyze the antiviral capabilities of LbuCas13a from Leptotrichia buccalis against various phages.
- To investigate the potential of LbuCas13a for broad-spectrum phage genome editing applications.
Main Methods:
- Challenging LbuCas13a against nine diverse E. coli phages to assess antiviral efficacy.
- Employing a two-step phage-editing and enrichment strategy for genome modification.
- Characterizing the efficiency and scope of LbuCas13a-mediated edits, including deletions and codon replacements.
Main Results:
- LbuCas13a exhibited robust antiviral activity, independent of target phage gene essentiality, expression timing, or location.
- Broad efficacy was confirmed against multiple E. coli phage groups.
- Achieved seven markerless genome edits in three distinct phages with 100% efficiency, including large deletions and single-codon changes.
Conclusions:
- LbuCas13a is a potent antiviral agent with broad-spectrum activity against diverse phages.
- This CRISPR-Cas13 system offers a versatile and efficient tool for precise phage genome editing.
- Cas13a represents a generalizable technology for manipulating the genomes of bacteriophages.
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