摘要
Saccharomyces cerevisiae is widely used as a model organism in biotechnology. However, its rigid cell wall limits efficient delivery of exogenous materials. In this study, we developed a nanoelectrode chip that integrated dielectrophoretic positioning, electrophoretic enrichment, and electroporation for yeast transformation. An accompanying data acquisition system enabled precise three-channel signal control for yeast capture, target concentration, and membrane permeabilization. MATLAB simulations showed that increasing the duty cycle accelerated plasmid DNA migration toward the nanoelectrodes. Experimentally, introducing the electrophoretic enrichment signal enhanced the local accumulation of charged dyes and plasmid DNA, whereas removal of enrichment signal significantly reduced transformation frequency. Using Kluyveromyces marxianus as an additional model organism, optimization of the duty cycle of the enrichment signal improved delivery frequency. Under electroporation conditions of 200 Hz and 10 Vpp, we achieved a fluorescence-based transformation frequency of 54.6 ± 3.8% and a yeast viability of 87.3 ± 8.5%, without cell wall removal or chemical pretreatment. Compared with conventional chemical methods, this approach offers shorter operation time, improved controllability, and potential for miniaturization and high-throughput applications. Overall, this platform provides a low-voltage and efficient strategy for fungal genetic engineering and single-yeast studies. © 2026 The Authors