DOI: https://doi.org/10.36347/sajb.2025.v13i07.003

Pages: 887-897

By providing previously unheard-of levels of accuracy, programmability, and efficiency, CRISPR-Cas systems have transformed genome editing and become potent instruments in biotechnology and biochemistry. The many mechanistic discoveries, new technical developments, and emerging uses of CRISPR-Cas technologies in molecular life sciences are examined in this review. The CRISPR-Cas immune defense system, which was first obtained from prokaryotes, functions mechanistically by recognizing and cleaving nucleic acids in a sequence-specific manner under the guidance of RNA molecules. Different Cas variations, such as Cas9, Cas12, and Cas13, exhibit distinct targeting and cleavage modalities. Our knowledge of guide RNA architecture, PAM sequence requirements, and conformational dynamics essential for substrate identification and editing fidelity has been expanded by structural and biochemical investigations. High-fidelity Cas variants, base editors, prime editors, and transposase-associated CRISPR tools are recent developments that broaden the editing toolset beyond basic double-strand breaks. These technological developments increase the range of editable genomic elements and reduce off-target consequences. Furthermore, new avenues for functional genomics and therapeutic applications have been made possible by the combination of CRISPR systems with synthetic biology, epigenome editing, diagnostics, and live-cell imaging. Microbial strain engineering, agricultural trait enhancement, environmental biosensing, and therapeutic genome repair in genetic disorders and immuno-oncology are examples of emerging uses. This study also emphasizes new delivery methods that promote safe and effective cellular absorption, such as ribonucleoprotein complexes, viral vectors, and nanoparticles. In order to put the social ramifications of CRISPR technology into perspective, the ethical, legal, and biosecurity issues pertaining to ecological gene drives and human germline editing are finally co