Literature DB >> 30826656

Portoporator©: A portable low-cost electroporation device for gene transfer to cultured cells in biotechnology, biomedical research and education.

Max A Schmitt1, Oliver Friedrich1, Daniel F Gilbert2.   

Abstract

Electroporation has been a widely established method for delivering DNA and other material into cells in vitro. Conventional electroporation infrastructure is typically immobile, non-customizable, non-transparent regarding the characteristics of output pulses, and expensive. Here, we describe a portable electroporator for DNA delivery into bacterial cells that can quickly be reconstructed using 3D desktop printing and off-the-shelf components. The device is light weight (700 g), small (70 × 180 × 210 mm) and extremely low-cost (<EUR 130). We provide the electrical circuitry and a detailed parts list for rebuilding the device. We characterize the properties of generated pulses and apply the system for gene delivery into bacterial Dh5α cells. We analyze the transformation efficiency based on the optical density of cell suspensions at 595 nm and on quantitative analysis of images obtained from bacterial cell-grown agar plates using colony forming units as well as confluence as indicators. We demonstrate time-dependency of the transformation efficiency using single pulses of 500 V between 1 and 1000 ms duration and we show that commercially available electroporation cuvettes of 1 mm gap size reveal higher transformation rates compared to cuvettes with 2 mm gap. We benchmark the transformation efficiency obtained using our platform with data from a heat shock-based transformation protocol and with data from a commercially available electroporator and show that our system reveals comparable results as the other techniques in the applied configurations. While this work focuses on genetic manipulation of bacterial cells, the device may also be applicable for delivery of genetic material small molecule or nanomaterials into other cell types, including mammalian cells.
Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Entities:  

Keywords:  3D printing; Arduino; Dh5α; Electroporation; Transformation

Mesh:

Substances:

Year:  2019        PMID: 30826656     DOI: 10.1016/j.bios.2019.02.024

Source DB:  PubMed          Journal:  Biosens Bioelectron        ISSN: 0956-5663            Impact factor:   10.618


  5 in total

1.  Modification of a Ready-Made High-Voltage Pulse Generator for Non-Thermal Irreversible Electroporation.

Authors:  K Zhang; J Wang
Journal:  Bull Exp Biol Med       Date:  2022-06-23       Impact factor: 0.804

2.  3D Printed Lab-on-a-Chip Platform for Chemical Stimulation and Parallel Analysis of Ion Channel Function.

Authors:  Daniel Aschenbrenner; Oliver Friedrich; Daniel F Gilbert
Journal:  Micromachines (Basel)       Date:  2019-08-19       Impact factor: 2.891

3.  Ultra-Low-Cost 3D Bioprinting: Modification and Application of an Off-the-Shelf Desktop 3D-Printer for Biofabrication.

Authors:  Melanie Kahl; Markus Gertig; Phillipp Hoyer; Oliver Friedrich; Daniel F Gilbert
Journal:  Front Bioeng Biotechnol       Date:  2019-07-31

Review 4.  Recent Advances in Stimulus-Responsive Nanocarriers for Gene Therapy.

Authors:  Cheng Yu; Long Li; Pei Hu; Yan Yang; Wei Wei; Xin Deng; Lu Wang; Franklin R Tay; Jingzhi Ma
Journal:  Adv Sci (Weinh)       Date:  2021-05-16       Impact factor: 16.806

5.  Mediated amperometry as a prospective method for the investigation of electroporation.

Authors:  Povilas Simonis; Rasa Garjonyte; Arunas Stirke
Journal:  Sci Rep       Date:  2020-11-05       Impact factor: 4.379

  5 in total

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