Banta Amy B, Cuellar Rodrigo A, Nadig Nischala, Davis Bryce C, Peters Jason M
Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA.
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.
Curr Opin Microbiol. 2025 Feb;83:102563. doi: 10.1016/j.mib.2024.102563. Epub 2024 Dec 3.
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.
CRISPR相关转座子(CASTs)是CRISPR-Cas机制与Tn7样转座子的天然融合体,可通过可编程的引导RNA指导转座子DNA的位点特异性整合。尽管基于CAST的转座机制在分子和结构层面已得到充分研究,但CASTs尚未广泛应用于细菌基因组工程和系统基因表型分析(即功能基因组学)——这可能是由于它们相对较新才被发现。在此,我们描述了CASTs的功能和应用,重点关注特征明确的系统,包括来自霍乱弧菌的I-F型CAST(VcCAST)和来自霍氏伪枝藻的V-K型CAST(ShCAST)。此外,我们通过提出对现有CAST工具进行基因组规模扩展的方案,探讨了靶向转座对细菌功能基因组学可能产生的变革性影响。