ADVANCEMENTS IN CRISPR-CAS9 TECHNOLOGY: REVOLUTIONIZING GENETIC EDITING AND THERAPEUTIC APPLICATIONS

Abstract

Molecular biology has seen the revolution of the CRISPR-Cas9 genome-editing mechanism with its accurate and efficient optional methodology in the targeted manipulation of the DNA. Initially based on the bacterial defense system, CRISPR-Cas9 has rapidly grown to become fundamental technology in therapeutic gene editing, functional genomics and agricultural biotechnology.We critically studied CRISPR-Cas9 applications in biomedical and agronomic settings, and more specifically analyzed aspects of CRISPR-Cas9 application efficiency, off-target activities, immunological responses and delivery mechanisms. Its methodology involved experimental testing, comparative analysis of the results of editing, and computational analysis of the frequencies of correcting mutations with and without HDR and NHEJ repair mechanisms. Columns and graphs were used in plotting results on gene disruption profile, delivery efficiencies and response pattern in various cell lines and under different experimental conditions.High editing rates of different cell lines were revealed, with base and prime editing variants having shown to be more specific. Other delivery methods including lipid nanoparticle and electroporation presented moderate to high efficiencies but off-target activity limits them, especially when high-fidelity variants of Cas9 are unavailable. In farming, gene-edited crops indicated huge improvement in yield and resistance features. Attempts to use Cas9 have also been found to elicit immune responses in some delivery settings, justifying the development of immune-evasive strategies.Summing up, CRISPR-Cas9 has been used to broaden the boundaries of genome engineering, but its overall potential in therapeutic and agronomic applications depends on the ability to resolve essential issues associated with off-target effects, delivery and regulatory control. Ethics, especially germline editing and equitable access, are still the major concerns in its ethical use. This study underlines the potential and the challenges of innovative CRISPR-driven technologies and requires communications through interdisciplinary strategies to make the potential advantages of such technology safe and inclusive.