Gene Editing for Tuberculosis using CRISPR-Cas Technology

Advances in genome editing technologies are simplifying the tedious, laborious work in research. Recent advancements have introduced innovative genome editing techniques, such as the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) and CRISPR-associated protein (Cas) system, which have proven to be powerful tools in genome editing. Unlike previous methods, CRISPR-Cas is highly specific and capable of gene knock-in, knock-out, or knockdown, which aids researchers. Such a tool is valuable for diagnosis, treatment, and vaccine production against life-threatening, mutating, and drug-resistant diseases such as Tuberculosis (TB). Mycobacterium Tuberculosis (Mtb) and other members of the Mycobacterium Tuberculosis Complex (MTBC) are notorious and slow-growing bacteria; hence, they are difficult to tackle. Different CRISPR-Cas systems, for example, Cas9, Cas12a, Cas12b, and Cas13a, are the types being used in the diagnosis of Mycobacterium tuberculosis. We have attempted to provide an overview of CRISPR-Cas technology and its application in tuberculosis. The literature was searched across platforms like PubMed, Google Scholar, Web of Science, SciELO, and Scopus. The keywords that were used to search the related literature were 'Mycobacterium tuberculosis OR Tuberculosis,' ‘TB AND Diagnosis,' ‘TB Vaccines AND Treatment,' ‘Mycobacterium tuberculosis AND CRISPR-Cas,' ‘CRISPR-Cas system,' ‘CRISPR-Cas in TB diagnosis,' 'Cas9,' 'Cas12a,' 'Cas13,' 'gene editing mycobacteria,' and ‘CRISPRi Mycobacterium tuberculosis.' The approach to searching for and selecting articles for the review provided a comprehensive and well-supported analysis of the subject matter. The review emphasizes the novel technology CRISPR-Cas system for diagnosis, treatment, and the development of new TB vaccines. Along with the applications, the review comprises traditional methods of diagnosis of TB and mechanisms and challenges related to CRISPR-Cas. Using the CRISPRi technology, new targets were found as promising novel drug targets. Vaccine development with the help of this tool is also mentioned in this review. CRISPR-Cas technology has emerged as a powerful tool for advancing TB research. Cas9, Cas12a, Cas12b, and Cas13 have demonstrated high specificity and sensitivity in detecting Mtb, even in samples with low bacterial loads. Moreover, CRISPR-mediated gene silencing has enhanced our understanding of the virulence mechanisms of the bacterium, facilitating the identification of novel therapeutic targets for designing new anti-TB drugs. Tuberculosis continues to pose a major global health challenge due to the persistence of the pathogen and the emergence of drug-resistant strains. CRISPR-Cas technology offers innovative solutions, and its applications extend beyond diagnostics to include drug discovery, therapeutic development, and vaccine research. Thus, CRISPR-Cas systems can significantly accelerate efforts toward effective control and eradication of tuberculosis.