Continuous cell electroporation for efficient DNA and siRNA delivery based on laminar microfluidic chips

Area of Science:

• Biotechnology
• Molecular Biology
• Bioengineering

Background:

• Electroporation is a widely used physical gene transfer method known for its high efficiency and low toxicity.
• Conventional electroporation techniques are typically restricted to small sample volumes, limiting throughput.
• There is a need for scalable and efficient gene delivery methods in biological research and therapeutic applications.

Purpose of the Study:

• To develop and present a continuous cell electroporation method utilizing microfluidic chip technology.
• To demonstrate the efficient delivery of genetic material (DNA and siRNA) into diverse cell lines using the developed system.
• To provide a detailed protocol for fabricating the microfluidic chip, setting up the apparatus, and performing the continuous electroporation assay.

Main Methods:

• Fabrication of a polydimethylsiloxane (PDMS) microfluidic chip using standard microfluidic chip fabrication techniques.
• Assembly of a continuous flow system using syringe pumps and a pulse generator.
• Optimization and execution of the continuous cell electroporation assay for gene delivery.

Main Results:

• Successful fabrication of a microfluidic chip suitable for continuous electroporation within 1-2 days.
• Demonstration of efficient delivery of both DNA and small interfering RNA (siRNA) into multiple cell lines.
• Establishment of a 1-hour protocol for the continuous electroporation assay, indicating high throughput potential.

Conclusions:

• The developed continuous cell electroporation method offers a scalable and efficient alternative to traditional batch electroporation.
• Microfluidic chip technology enables the fabrication of user-friendly devices for high-throughput gene transfer.
• This method holds promise for applications in cell biology research, drug screening, and potentially gene therapy.