Utilizing Digital Microfluidics to Miniaturize and Automate Arrayed CRISPR Screening Workflows

Drug developers are consistently aiming to enhance clinical relevance in drug discovery. CRISPR has emerged as a versatile tool to accelerate the entire drug discovery process, with arrayed CRISPR screening offering extensive genome coverage and richer datasets. However, conducting arrayed CRISPR screens with translatable cell models (such as primary cells, iPSCs, etc.) presents challenges due to cost and the need for large cellular inputs, which often necessitates donor pooling and results in heterogeneous datasets. To address these challenges and facilitate genome-wide arrayed CRISPR screening in patient-derived cells, a novel automated microfluidic system has been developed. This system miniaturizes non-viral transfection, significantly reducing the number of cells required per edit while maintaining high efficiency and viability. It enables efficient delivery of various payloads (mRNA, DNA, proteins, RNP) and gene editing across different cell types and structures (including primary immune cells, iPSCs). A custom 48-plex editing cartridge and its associated workflow for automation integration have been introduced, allowing for thousands of edits daily without human intervention. This approach offers the potential for rapid, cost-effective, high-throughput discovery at the resolution of individual patients, which could lead to a reduction in drug candidate attrition and the advancement of personalized therapies.

Learning Objectives: 

1. Demonstrate Knowledge with gene editing, microfluidics and non-viral delivery methods.

2. Review the uses of a low-volume high-throughput transfection platform for gene editing applications.

3. Discover the applicability of an automation-ready digital microfluidics platform for high-throughput arrayed CRISPR screening.