The promise of CRISPR-associated transposons for bacterial functional genomics.

Curr Opin Microbiol

Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA; Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, WI 53726, USA; Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA; Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, USA; Center for Genomic Science Innovation, University of Wisconsin-Madison, Madison, WI 53706, USA. Electronic address:

Published: December 2024

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.

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Source
http://dx.doi.org/10.1016/j.mib.2024.102563DOI Listing

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