Novel electroporation microchip with field constriction enhances transfection efficiency and survival rates of feline embryos

Abstract

This study introduces a low-voltage electroporation microchip designed for transfection in cat embryos, featuring real-time impedance monitoring. The microchip uses a field constriction strategy, which localises the electric field to the membrane region in contact with the micro-orifice, enhancing electroporation efficiency while minimising damage. Embryos were positioned on the orifice, and a series of voltage pulses (10, 15, and 20 V) were applied. Electroporation efficacy was assessed using fluorescent dyes, followed by real-time impedance measurements to monitor the membrane resealing time. It provided valuable insights into membrane recovery times, essential for optimizing gene editing conditions to ensure efficient delivery and maintain cell integrity. The results demonstrated that the microchip with 15 V achieved a 69.5% higher electroporation rate and 100% of survival compared to conventional devices (p < 0.05). Additionally, the microchip successfully facilitated the transfer of green fluorescent protein genes into embryos, achieving a 78.5% success rate significantly greater compared to 53.6% with the conventional device (p < 0.05). This innovative microchip provides transformative transfection technology for safer and more efficient genomic modifications in embryos. It holds promising applications across species and therapeutic interventions, paving the way for future studies in advanced genomic research.