Jul 2026· Open Microbiology Journal· 0 citations· 100 references
TL;DR
Traditional methods of diagnosis of TB and mechanisms and challenges related to CRISPR-Cas are included, including the novel technology CRISPR-Cas system for diagnosis, treatment, and the development of new TB vaccines.
Abstract
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.
The potential application of CRISPR technology for the possible management of geneticbased conditions, including sickle-cell anemia, β-thalassemia, cystic fibrosis, and Duchenne muscular dystrophy is described.
M. Veer, Poonam Nikam, Omkar More et al.· International Journal of Dru...· 0 citations
Over a decade of advances in Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR) and CRISPR-associated protein 9 (Cas9)-based technologies have culminated in the first-ever FDA-approved CRISPR/Cas-based therapy. Aside from this approved therapy for sickle cell anemia, several CRISPR/Cas-based therapies are currently under development or testing for a range of chronic diseases, including viral diseases like human immunodeficiency virus type 1 (HIV-1) infection, genetic diseases like familial hypercholesterolemia, and cancer. The success of these therapies hinges on the effective delivery of CRISPR/Cas9 components to target regions, efficient Cas endonuclease editing, repair profiles generated, and their resulting outcomes. Here, we discuss the factors that influence the generation of CRISPR/Cas9-generated repair edits, the overall profiles, and outcome prediction(s), as well as the analytical tools that have been developed to date. Finally, how this technology has been used towards a functional HIV-1 cure is discussed.
Samuel N. Effah, Shirley C. Barrera, Nahia Urturi Ortiz et al.· International Journal of Mol...· 0 citations
The application of genome editing, CRISPR/Cas9 has revolutionized plant breeding by enabling precise, efficient, and targeted modification of native genes, significantly accelerating the development of improved agronomic traits of crops. Therefore, CRISPR/Cas9 technology currently the most extensively used genome editing technique worldwide because of its simple design, cost-effectiveness, high efficiency, good reproducibility, high engineering feasibility, ability to create gene knockout, RNA editing, and quick cycle. It is used to knock in or knock out genes of interest and for generating models for genetic studies. The main components of the CRISPR/Cas9 system are an RNA-guided Cas9 endonuclease and a single-guide RNA (sgRNA). The workflow of CRISPR/Cas9 gene editing comprises selecting target sites, designing and synthesizing sgRNA, introducing transformation constructs or ribonucleoprotein (RNP) in plant cells, followed by transformation and identification of edited lines. This approach bypasses the formal regulations on GMOs, thus encouraging the widespread adoption RNA-guided gene editing in agricultural sciences and biotechnology. The system is now being utilized in the biofortification of cereal crops such as rice, wheat, barley, and maize, including vegetable crops such as potato and tomato. The world's first genome-edited rice varieties are DRR Dhan 100 (Kamala) and Pusa DST Rice 1 developed by the Indian Council of Agricultural Research (ICAR), New Delhi, India in 2025 with the objective of bringing about revolutionary changes in terms of higher production, climate adaptability, and water conservation. The CRISPR/Cas9-based crop genome editing has been utilized in imparting/producing qualitative enhancement in aroma, shelf life, sweetness, and quantitative improvement in starch, protein, gamma-aminobutyric acid (GABA), oleic acid, anthocyanin, phytic acid, gluten, and steroidal glycoalkaloid contents. Some varieties have even been modified to become disease and stress-resistant. Therefore, CRISPR/Cas9 is aiding in developing climate-ready crops and improving crop quality parameters such as appearance, palatability, nutritional components, and other preferred traits. Gene editing tools are used to generate changes to the native genetic material. Unlike GMOs, which introduce novel configurations of genetic materials typically derived from other organisms, gene editing methods modify existing genetic material in ways that can yield beneficial outcomes.
Ravindra B. Malabadi, Raju K. Chalannavar· World Journal of Advanced Re...· 0 citations
This review systematically summarizes the developmental logic, core mechanisms, clinical applications, advantages and limitations of the three generations of CRISPR-Cas technology in monogenic disorders, and analyzes the key challenges such as delivery efficiency, long-term safety, and treatment accessibility.
Yiwen Wang· Theoretical and Natural Scie...· 0 citations