The promise of CRISPR-associated transposons for bacterial functional genomics

Amy B Banta; Rodrigo A Cuellar; Nischala Nadig; Bryce C Davis; Jason M Peters

doi:10.1016/j.mib.2024.102563

The promise of CRISPR-associated transposons for bacterial functional genomics

Curr Opin Microbiol. 2024 Dec 3:83:102563. doi: 10.1016/j.mib.2024.102563. Online ahead of print.

Authors

Amy B Banta¹, Rodrigo A Cuellar², Nischala Nadig³, Bryce C Davis¹, Jason M Peters⁴

Affiliations

¹ Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA.
² Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA; Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, WI 53706, USA; Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, WI 53726, USA.
³ Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA; Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, WI 53706, USA.
⁴ 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: [email protected].

PMID: 39631148
DOI: 10.1016/j.mib.2024.102563

Abstract

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.

Publication types

Review