A Comparative Analysis of CRISPR-Cas9 and Base Editing Approaches for Precise Genomic Modifications in Eukaryotic Systems

Abstract

The advent of genome editing technologies has catalyzed transformative advancements in molecular biology, with CRISPR-Cas9 and base editing emerging as front-runners for precise genomic modifications. This study employs a multifaceted evaluation of these two methodologies, assessing their efficiency, accuracy, and off-target effects through a series of in vitro experiments utilizing HEK293T cells. We utilized advanced sequencing techniques, including Sanger and Next-Generation Sequencing (NGS), to quantify modification rates and identify potential off-target loci. Our findings reveal that while CRISPR-Cas9 provides robust editing capabilities, base editing significantly minimizes off-target effects, resulting in a 30% decrease in unintended modifications. These results elucidate the strengths and limitations of each method, providing a comprehensive framework for researchers to inform their choices in genomic editing applications. This study not only contributes to the existing literature by directly comparing these techniques but also emphasizes the critical need for tailored approaches in diverse genomic contexts for future applications in gene therapy and synthetic biology.