An automated microfluidic gene-editing platform for deciphering cancer genes

Area of Science:

• Biotechnology
• Molecular Biology
• Microfluidics

Background:

• Gene editing, particularly RNA-guided endonuclease systems, is vital for phenotypic screening.
• Existing methods for gene editing assays are often arrayed or pooled, prompting a need for new technologies.
• Automating gene editing assays can accelerate research and drug discovery.

Purpose of the Study:

• To develop and validate the first digital microfluidic method for automating arrayed gene editing in mammalian cells.
• To demonstrate the platform's utility in culturing lung cancer cells and performing gene knockout.
• To assess the platform's application in drug sensitivity testing for personalized medicine.

Main Methods:

• Development of a digital microfluidic system for automated cell culture, gene transfection, and knockout.
• Implementation of a standardized imaging pipeline for analyzing fluorescently labeled cells.
• Conducting a gene editing assay targeting the MAPK/ERK pathway in lung cancer cells, including RAF1 gene knockout.

Main Results:

• The digital microfluidic platform successfully cultured lung cancer cells for up to six days.
• Automated gene transfection and knockout procedures were efficiently implemented.
• Combined gene editing and drug treatment (Sorafenib Tosylate) on the device reduced the half-inhibitory concentration (IC50) compared to drug treatment alone.

Conclusions:

• The developed digital microfluidic system enables automated arrayed gene editing in mammalian cells.
• This platform is effective for gene knockout and drug sensitivity assays in lung cancer cells.
• The system holds significant potential for advancing gene-editing assays and personalized medicine.