Advances in CRISPR-Cas9 gene editing: Biochemical applications and future directions

AYESA, BIBI and Begum, Mst. Shahinur and Iqbal, Farheen and Urmi, Banasree Roy and Malakar, Rajib Kumar and Hossain, Sayed and Rashed, Md. Jobaer Rahman (2025) Advances in CRISPR-Cas9 gene editing: Biochemical applications and future directions. International Journal of Science and Research Archive, 15 (3). pp. 1771-1782. ISSN 2582-8185

Because of CRISPR-Cas9 technology, genome engineering and molecular biology are evolving in new directions. Borrowing from the immune systems of early lifeforms, the CRISPR-Cas9 technology is used to rearrange genetic codes in many organisms precisely. It covers recent developments in CRISPR-Cas9 and highlights its biochemical functions and more uses for this technology. We look into how the structure of the CRISPR-Cas9 complex contributes to guide RNA (gRNA), the use of PAM sequences, and variants of Cas9 such as dCas9, base editors, and prime editors. Recently, RNA editing, epigenetic reprogramming, and diagnostics using CRISPR have been shown to have far more uses than just changing genes. In this area, CRISPR-Cas9 helps to better understand the workings of genes, enhance how metabolic processes function, and change microorganisms for business uses. Using it to help with monogenic conditions, modify the immune system, and deal with infectious diseases has seen fast progress in moving from the lab to patient care. Among the important points in the review are limits like off-target action, difficulty delivering the therapies, and immunogenicity, which continue to block the wide-scale use of gene therapies in medicine. In the last section, the paper outlines what lies ahead, such as using artificial intelligence to guide RNA design, designing CRISPR-based circuits for synthetic biology, and considering the ethics involved with germline editing. Bringing together recent studies, this article points out that CRISPR-Cas9 is becoming increasingly important as a platform for new biotechnology and medicine.