High-Throughput Microfluidic Electroporation (HTME): A Scalable, 384-Well Platform for Multiplexed Cell Engineering.
| Title: | High-Throughput Microfluidic Electroporation (HTME): A Scalable, 384-Well Platform for Multiplexed Cell Engineering. |
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| Authors: | Gaillard WR; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Sandia National Laboratories, Livermore, CA 94550, USA.; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Sustarich J; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Sandia National Laboratories, Livermore, CA 94550, USA.; Li Y; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Sandia National Laboratories, Livermore, CA 94550, USA.; Engineering Directorate, Lawrence Livermore National Laboratory, Livermore, CA 94550, USA.; Carruthers DN; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Gupta K; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Engineering Directorate, Lawrence Livermore National Laboratory, Livermore, CA 94550, USA.; Liang Y; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Kuo R; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Tan S; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Yoder S; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Adams PD; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Department of Bioengineering, University of California Berkeley, Berkeley, CA 94720, USA.; Garcia Martin H; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; BCAM-Basque Center for Applied Mathematics, 48009 Bilbao, Spain.; Hillson NJ; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Singh AK; DOE Joint BioEnergy Institute, Emeryville, CA 94608, USA.; Sandia National Laboratories, Livermore, CA 94550, USA.; Engineering Directorate, Lawrence Livermore National Laboratory, Livermore, CA 94550, USA. |
| Source: | Bioengineering (Basel, Switzerland) [Bioengineering (Basel)] 2025 Jul 22; Vol. 12 (8). Date of Electronic Publication: 2025 Jul 22. |
| Publication Type: | Journal Article |
| Language: | English |
| Journal Info: | Publisher: MDPI AG Country of Publication: Switzerland NLM ID: 101676056 Publication Model: Electronic Cited Medium: Print ISSN: 2306-5354 (Print) Linking ISSN: 23065354 NLM ISO Abbreviation: Bioengineering (Basel) Subsets: PubMed not MEDLINE |
| Imprint Name(s): | Original Publication: Basel, Switzerland : MDPI AG, [2014]- |
| Abstract: | 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. |
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| Grant Information: | DE-AC02-05CH11231 United States Department of Energy |
| Contributed Indexing: | Keywords: automation; electroporation; high-throughput; microfluidic; self-driving lab; strain engineering; synthetic biology; transfection; transformation |
| Entry Date(s): | Date Created: 20250828 Date Completed: 20250914 Latest Revision: 20250914 |
| Update Code: | 20260130 |
| PubMed Central ID: | PMC12383916 |
| DOI: | 10.3390/bioengineering12080788 |
| PMID: | 40868301 |
| Database: | MEDLINE |
Journal Article