High-Throughput Microfluidic Electroporation (HTME): A Scalable, 384-Well Platform for Multiplexed Cell Engineering

Area of Science:

• Synthetic Biology
• Biotechnology
• Molecular Biology

Background:

• Electroporation is key for gene delivery in synthetic biology but faces scalability and tunability challenges in multiplexed workflows.
• Current methods are costly and time-consuming for screening genetic libraries and optimizing experimental parameters.
• Limitations hinder rapid Design-Build-Test-Learn (DBTL) cycles in synthetic biology.

Purpose of the Study:

• To develop a High-Throughput Microfluidic Electroporation (HTME) platform to overcome limitations in current electroporation techniques.
• To enable rapid, cost-effective, and scalable gene delivery for synthetic biology applications.
• To facilitate the optimization of electroporation parameters and accelerate DBTL cycles.

Main Methods:

• Developed a High-Throughput Microfluidic Electroporation (HTME) platform featuring a 384-well electroporation plate (E-Plate) and control electronics.
• Utilized cost-effective printed-circuit-board (PCB) technology for E-Plate fabrication, enabling nano to microliter volumes.
• Implemented individual well control for rapid optimization and integration into automated workflows.

Main Results:

• The HTME platform can electroporate all 384 wells in under a minute with individual control.
• Demonstrated successful transformation of E. coli with pUC19, achieving >99% well success rate for single colony forming units.
• Validated the platform's capability for rapid, customizable electroporations across hundreds of conditions.

Conclusions:

• The HTME platform significantly reduces time and cost associated with gene delivery in synthetic biology.
• This technology addresses the bottleneck in transformation/transfection, accelerating DBTL cycles.
• The HTME platform offers a scalable and tunable solution for high-throughput genetic engineering.