| Title: |
High-Throughput Microfluidic Electroporation (HTME): A Scalable, 384-Well Platform for Multiplexed Cell Engineering |
| Authors: |
Gaillard, William R; Sustarich, Jess; Li, Yuerong; Carruthers, David N; Gupta, Kshitiz; Liang, Yan; Kuo, Rita; Tan, Stephen; Yoder, Sam; Adams, Paul D; Martin, Hector Garcia; Hillson, Nathan J; Singh, Anup K |
| Source: |
Bioengineering, vol 12, iss 8 |
| Publisher Information: |
eScholarship, University of California |
| Publication Year: |
2025 |
| Collection: |
University of California: eScholarship |
| Subject Terms: |
40 Engineering (for-2020); 4003 Biomedical Engineering (for-2020); Biotechnology (rcdc); Genetics (rcdc); Bioengineering (rcdc); Generic health relevance (hrcs-hc); 12 Responsible Consumption and Production (sdg); high-throughput; electroporation; transformation; transfection; automation; microfluidic; self-driving lab; synthetic biology; strain engineering |
| Subject Geographic: |
788 - 788 |
| Description: |
Electroporation-mediated gene delivery is a cornerstone of synthetic biology, offering several advantages over other methods: higher efficiencies, broader applicability, and simpler sample preparation. Yet, electroporation protocols are often challenging to integrate into highly multiplexed workflows, owing to limitations in their scalability and tunability. These challenges ultimately increase the time and cost per transformation. As a result, rapidly screening genetic libraries, exploring combinatorial designs, or optimizing electroporation parameters requires extensive iterations, consuming large quantities of expensive custom-made DNA and cell lines or primary cells. To address these limitations, we have developed a High-Throughput Microfluidic Electroporation (HTME) platform that includes a 384-well electroporation plate (E-Plate) and control electronics capable of rapidly electroporating all wells in under a minute with individual control of each well. Fabricated using scalable and cost-effective printed-circuit-board (PCB) technology, the E-Plate significantly reduces consumable costs and reagent consumption by operating on nano to microliter volumes. Furthermore, individually addressable wells facilitate rapid exploration of large sets of experimental conditions to optimize electroporation for different cell types and plasmid concentrations/types. Use of the standard 384-well footprint makes the platform easily integrable into automated workflows, thereby enabling end-to-end automation. We demonstrate transformation of E. coli with pUC19 to validate the HTME's core functionality, achieving at least a single colony forming unit in more than 99% of wells and confirming the platform's ability to rapidly perform hundreds of electroporations with customizable conditions. This work highlights the HTME's potential to significantly accelerate synthetic biology Design-Build-Test-Learn (DBTL) cycles by mitigating the transformation/transfection bottleneck. |
| Document Type: |
article in journal/newspaper |
| File Description: |
application/pdf |
| Language: |
unknown |
| Relation: |
qt5xf4s2kv; https://escholarship.org/uc/item/5xf4s2kv; https://escholarship.org/content/qt5xf4s2kv/qt5xf4s2kv.pdf |
| DOI: |
10.3390/bioengineering12080788 |
| Availability: |
https://escholarship.org/uc/item/5xf4s2kv; https://escholarship.org/content/qt5xf4s2kv/qt5xf4s2kv.pdf; https://doi.org/10.3390/bioengineering12080788 |
| Rights: |
public |
| Accession Number: |
edsbas.DB5A0F5E |
| Database: |
BASE |