ENGINEERING NOVEL CRISPR PLATFORMS FOR HIGH-FIDELITY SOMATIC CELL GENE CORRECTION

Abstract

Background: The emergence of CRISPR-based gene editing has revolutionized therapeutic gene correction through precise editing of disease-causing mutations. However, out of bounds mutations, genetic instability, double-strand DNA break toxicity, and shipping inefficiencies still pose major obstacles for safe somatic cell applications.

Objective: The study examines the current state of engineering approaches for developing high-fidelity CRISPR structures to correct somatic genes efficiently and precisely while minimizing off-target activity.

Method: Here, we conducted an evaluation of engineered Cas variations, optimized guide RNAs, base modification systems, prime editing technologies and specific distribution platforms to summarize current translational genome engineering research.

Results: High-fidelity CRISPR variants showed substantially decreased off-target cleavage but retained efficient on-target editing. Engineered base editors resulted in ~75-92% correction efficiency in somatic cell models with lower double strand break toxicity. Prime editing systems increased editing precision and decreased insertion-deletions. In addition, optimized lipid nanoparticle and viral delivery systems greatly enhanced intracellular editing efficiency, cell viability, and therapeutic gene restoration in corrected somatic cells.

Conclusion: Engineering next generation high fidelity CRISPR platforms provides a transformative approach for safe and precise somatic gene correction. However, long-term genomic safety, delivery optimization, immunogenicity reduction and clinical scalability remain important challenges for future therapeutic application.