Research Progress of CRISPR-Cas3

The CRISPR-Cas system has revolutionized genome editing. Among its nucleases, Cas9 and Cas12 have been studied most extensively. However, CRISPR-Cas3 stands out as a distinct nuclease, possessing both helicase and exonuclease activities. Despite this unique feature, it remains under-explored—especially considering its potential for large-scale genome manipulation. This review emphasizes the challenge of multidrug-resistant bacteria in current antimicrobial therapy, where Cas3 may provide a breakthrough solution by degrading the entire pathogen genome, rather than targeting single genes. To compile this review, a comprehensive literature search was conducted, covering publications from 2018 to 2025 across databases such as PubMed, Web of Science, and Nature. A comparative analysis of Cas3 and other CRISPR systems was also carried out to identify key research gaps. Key findings highlight Cas3's ability to degrade long DNA segments, making it valuable for deleting large genomic regions, such as integrated viral DNA, and addressing challenges in antimicrobial therapy. Nevertheless, low targeting precision and complex delivery processes hinder its clinical translation. The review concludes that to unlock Cas3's full potential, two steps are essential: optimizing its specificity through protein engineering and developing targeted delivery systems.

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