Broad-spectrum CRISPR-Cas13a enables efficient phage genome editing.
Nat Microbiol
; 7(12): 1967-1979, 2022 12.
Article
en En
| MEDLINE
| ID: mdl-36316451
ABSTRACT
CRISPR-Cas13 proteins are RNA-guided RNA nucleases that defend against incoming RNA and DNA phages by binding to complementary target phage transcripts followed by general, non-specific RNA degradation. Here we analysed the defensive capabilities of LbuCas13a from Leptotrichia buccalis and found it to have robust antiviral activity unaffected by target phage gene essentiality, gene expression timing or target sequence location. Furthermore, we find LbuCas13a antiviral activity to be broadly effective against a wide range of phages by challenging LbuCas13a against nine E. coli phages from diverse phylogenetic groups. Leveraging the versatility and potency enabled by LbuCas13a targeting, we applied LbuCas13a towards broad-spectrum phage editing. Using a two-step phage-editing and enrichment method, we achieved seven markerless genome edits in three diverse phages with 100% efficiency, including edits as large as multi-gene deletions and as small as replacing a single codon. Cas13a can be applied as a generalizable tool for editing the most abundant and diverse biological entities on Earth.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Bacteriófagos
/
Edición Génica
Idioma:
En
Revista:
Nat Microbiol
Año:
2022
Tipo del documento:
Article
País de afiliación:
Estados Unidos