A review: CRISPR/Cas12-Mediated Genome Editing in Fungal Cells: Advancements, Mechanisms, and Future Directions in Plant-Fungal Pathology

Abstract

The CRISPR-associated protein (CRISPR/Cas) system, characterized by clustered regularly interspaced short palindromic repeats, has revolutionized life science research by providing vast possibilities for alteringspecific DNA or RNA sequences in a variety of organisms. The present system integrates fragments of exogenous DNA, known as spacers, into CRISPR cassettes. These cassettes are subsequently transcribed into CRISPR arrays, which are further processed to generate guide RNA (gRNA). The CRISPR arrays are genetic loci that are responsible for encoding Cas proteins. The Cas proteins are responsible for supplying the necessary enzymatic machinery to acquire new spacers that are aimed at invading elements. The development of novel genome engineering tools has been made possible by utilizing various Cas proteins, including but not limited to Cas9, Cas12, Cas13, and Cas14, which possess programmable sequence specificity. The emergence of Cas variants has spurred genetic research and advanced the utilization of the CRISPR/Cas tool to manipulate and edit nucleic acid sequences within a wide range of living organisms. This review aims to furnishoperational modalities ofthe Cas12 protein identified thus far. Furthermore, the advantages and disadvantages ofCas12proteinare examined, along with their recent implementations in the plant fungal world. This review provides the researcher with insights into diverse Cas proteins and their potential applications in advanced genome editing for the next generation.