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.

Details

Title
Novel electroporation microchip with field constriction enhances transfection efficiency and survival rates of feline embryos
Author
Kallayanathum, Wirakan 1 ; Techaumnat, Boonchai 2 ; Panklang, Nitipong 3 ; Sripumkhai, Witsaroot 4 ; Jeamsaksiri, Wutthinan 4 ; Koo, Okjae 5 ; Jang, Goo 6 ; Tharasanit, Theerawat 1 

 Chulalongkorn University, Department of Obstetrics, Gynaecology, and Reproduction, Faculty of Veterinary Science, Bangkok, Thailand (GRID:grid.7922.e) (ISNI:0000 0001 0244 7875); Chulalongkorn University, Center of Excellence for Veterinary Clinical Stem Cells and Bioengineering, Bangkok, Thailand (GRID:grid.7922.e) (ISNI:0000 0001 0244 7875) 
 Chulalongkorn University, Department of Electrical Engineering, Faculty of Engineering, Bangkok, Thailand (GRID:grid.7922.e) (ISNI:0000 0001 0244 7875); Chulalongkorn University, Micro/Nano-Electro-Mechanical Integrated System Research Unit, Faculty of Engineering, Bangkok, Thailand (GRID:grid.7922.e) (ISNI:0000 0001 0244 7875) 
 Rajamangala University of Technology Thanyaburi, Department of Electrical Engineering, Faculty of Engineering, Pathum Thani, Thailand (GRID:grid.440403.7) (ISNI:0000 0004 0646 5810) 
 National Science and Technology Development Agency (NSTDA), Thai Microelectronics Center (TMEC), Chachoengsao, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408) 
 Kyungpook National University, College of Veterinary Medicine, Daegu, Republic of Korea (GRID:grid.258803.4) (ISNI:0000 0001 0661 1556); nSAGE Inc., Incheon, Republic of Korea (GRID:grid.258803.4) 
 Seoul National University, Department of Veterinary Clinical Science, College of Veterinary Medicine and the Research Institute of Veterinary Science, Seoul, Republic of Korea (GRID:grid.31501.36) (ISNI:0000 0004 0470 5905) 
Pages
29508
Publication year
2024
Publication date
2024
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
3133634871
Copyright
© The Author(s) 2024. This work is published under http://creativecommons.org/licenses/by-nc-nd/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.