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

摘要

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 x 180 x 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 Dh5a 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.