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Keywords: CRISPR, Gene-editing, CRISPR-Cas12a, Knock-in, Knock-out
Traditional methods for plant genetic transformation predominantly involve random DNA integration. However, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas techniques have emerged from an RNA-guided adaptive defense mechanism utilized by bacteria and archaea to protect against plasmids and phages. The CRISPR-Cas toolkit, a genome-editing tool for crop improvement, promises to accelerate plant breeding, reshape crop production, and revolutionize disease management. Various classes of CRISPR-Cas systems have been identified, and detailed insights into these different classes for developing future crops have been reported. CRISPR-Cas9 is the most used system so far. Cas12a (Cpf1), an RNA-guided endonuclease associated with the CRISPR adaptive immune system found in many prokaryotes, stands out for its unique ability to recognize A/T-rich DNA sequences and directly process its guide RNA, unlike its more prominent counterpart, Cas9. Applications of CRISPR-Cas systems in agriculture include improving crop traits, detecting and managing plant diseases, inactivating plant DNA viruses, improving nematode resistance, combating multidrug-resistant (MDR) bacterial infections, mitigating plant stress, enhancing climate resilience, overcoming abiotic and biotic stress conditions, and nutrient biofortification in crop plants (Kumar et al. in Kumar, Arora, Ogita, Yau, Mukherjee (eds) Gene editing in plants, Springer, Singapore, 2024c). This article will showcase selected cases of current applications and recent achievements in utilizing CRISPR-Cas9 and CRISPR-12a systems for enhancing crop improvement.
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Department of Botany and Biotechnology, University of Rajasthan, Jaipur, India