Microelectroporation device for genomic screening

摘要

We have developed an microelectroporation device that combines microarrays of oligonucleotides, microfluidic channels, and electroporation for cell transfection and high-throughput screening applications (e.g. RNA interference screens). Microarrays allow the deposition of thousands of different oligonucleotides in microscopic spots. Microfluidic channels and microwells enable efficient loading of cells into the device and prevent cross-contamination between different oligonucleotides spots. Electroporation allows optimal transfection of nucleic acids into cells (especially hard-to-transfect cells such as primary cells) by minimizing cell death while maximizing transfection efficiency. This invention has the advantage of a higher throughput and lower cost, while preventing cross-contamination compared to conventional screening technologies. Moreover, this device does not require bulky robotic liquid handling equipment and is inherently safer given that it is a closed system.

主权项

1. A microelectroporation device, comprising:
an optically transparent substrate; a printed circuit board (PCB); a biocompatible gasket disposed between and fluidically sealing a bottom surface of the PCB to a top surface of the optically transparent substrate; and a low voltage power supply;wherein the optically transparent substrate comprises a networked plurality of microfluidic channels and one or more arrays of microchambers formed into the top surface of the optically transparent substrate, and a conductive metal oxide layer deposited over the top surface to provide a common electrode,wherein the PCB comprises an array of discrete, addressable microelectrodes disposed on the bottom surface of the PCB configured and patterned to align with each of the one or more arrays of microchambers to provide a single microelectrode adjacent to each one of the microchambers, and wherein a thickness of the PCB is removed from surface comprising the array of addressable microelectrodes around each of the microelectrode in order to provide clearance for the biocompatible gasket; andwherein the biocompatible gasket comprises an array of through-holes configured to match both the array of microelectrodes and the one or more arrays of microchambers thereby providing for fluid and electrical communication between each one of the microchamber and the single microelectrode adjacent thereto.