Droplets for Gene Editing Using CRISPR-Cas9 and Clonal Selection Improvement Using Hydrogels

Abstract

Gene editing tools have triggered a revolutionary transformation in the realms of cellularand molecular physiology, serving as a fundamental cornerstone for the evolution of disease modelsand assays in cell culture reactions, marked by various enhancements. Concurrently, microfluidicshas emerged over recent decades as a versatile technology capable of elevating performance andreducing costs in daily experiments across diverse scientific disciplines, with a pronounced impacton cell biology. The amalgamation of these groundbreaking techniques holds the potential to amplifythe generation of stable cell lines and the production of extracellular matrix hydrogels. Thesehydrogels, assuming a pivotal role in isolating cells at the single-cell level, facilitate a myriad ofanalyses. This study presents a novel method that seamlessly integrates CRISPR-Cas9 gene editingtechniques with single-cell isolation methods in induced pluripotent stem cell (hiPSC) lines, utilizingthe combined power of droplets and hydrogels. This innovative approach is designed to optimizeclonal selection, thereby concurrently reducing costs and the time required for generating a stablegenetically modified cell line. By bridging the advancements in gene editing and microfluidictechnologies, our approach not only holds significant promise for the development of disease modelsand assays but also addresses the crucial need for efficient single-cell isolation. This integrationcontributes to streamlining processes, making it a transformative method with implications forenhancing the efficiency and cost-effectiveness of stable cell line generation. As we navigate theintersection of gene editing and microfluidics, our study marks a significant stride toward innovativemethodologies in the dynamic landscape of cellular and molecular physiology research.