The promise of CRISPR-associated transposons for bacterial functional genomics

CRISPR-associated transposons (CASTs) are naturally occurring amalgamations of CRISPR-Cas machinery and Tn7-like transposons that direct site-specific integration of transposon DNA via programmable guide RNAs. Although the mechanisms of CAST-based transposition have been well studied at the molecula...

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Bibliographic Details
Published in:Current opinion in microbiology 2025-02, Vol.83 (C), Article 102563
Main Authors: Banta, Amy B, Cuellar, Rodrigo A, Nadig, Nischala, Davis, Bryce C, Peters, Jason M
Format: Article
Language:English
Online Access:Get full text
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Summary:CRISPR-associated transposons (CASTs) are naturally occurring amalgamations of CRISPR-Cas machinery and Tn7-like transposons that direct site-specific integration of transposon DNA via programmable guide RNAs. Although the mechanisms of CAST-based transposition have been well studied at the molecular and structural level, CASTs have yet to be broadly applied to bacterial genome engineering and systematic gene phenotyping (i.e. functional genomics) — likely due to their relatively recent discovery. Here, we describe the function and applications of CASTs, focusing on well-characterized systems, including the type I-F CAST from Vibrio cholerae (VcCAST) and type V-K CAST from Scytonema hofmanni (ShCAST). Further, we discuss the potentially transformative impact of targeted transposition on bacterial functional genomics by proposing genome-scale extensions of existing CAST tools.
ISSN:1369-5274
DOI:10.1016/j.mib.2024.102563