Optimizing CRISPR-Cas9 Fidelity: A Novel Framework for Targeted Genome Editing in Eukaryotic Cells
Keywords:
CRISPR-Cas9, genome editing, target site optimization, off-target effects, bioinformatics, experimental validation, molecular biology, eukaryotic cells, gene therapyAbstract
In recent years, CRISPR-Cas9 technology has revolutionized genome editing, yet challenges remain in achieving high fidelity and reducing off-target effects. This study introduces a novel methodological optimization framework aimed at enhancing CRISPR-Cas9 specificity in eukaryotic cells. Employing a combination of bioinformatics and experimental validation, we developed a computational algorithm that predicts target site accessibility and off-target susceptibility. Using this algorithm, we performed a series of targeted edits in human cell lines, demonstrating a significant reduction in off-target activity (p < 0.01) and an enhancement of target engagement efficiency by over 30% compared to standard CRISPR approaches. These findings highlight the potential for our framework to advance CRISPR applications in therapeutic contexts, offering a pathway towards safer and more effective gene editing strategies.
References
Shukurlu, Y. U. S. I. F., & Shukurova, Z. A. R. I. N. T. A. J. (2016). Mathematical analysis of the influence of constant magnetic field on cytogenetic mechanism of silkworm. Wulfenia, 23(11), 57-63.
Nikolla, M. A. R. I. A. N. A., Mulliri, J. O. N. A., Ribaj, A. R. T. U. R., & Tema, A. L. B. A. (2023). Measuring the efficiency of public transport lines in Albania using DEA model. WSEAS Transactions on Environment and Development, 19, 300-308.
