Microfluidic platforms for RNA interference screening of virus-host interactions.

Area of Science:

• Genomics
• Virology
• Microfluidics

Background:

• RNA interference (RNAi) is crucial for functional genomics and analyzing host-pathogen interactions.
• Current high-throughput RNAi screening methods are often expensive, tedious, and require specialized equipment.
• There is a need for more accessible and cost-effective RNAi screening platforms.

Purpose of the Study:

• To develop an inexpensive, miniaturized microfluidic system for RNA interference (RNAi) screening.
• To enable on-chip patterning and screening of gene targets involved in virus entry and infection pathways.
• To reduce reagent usage and eliminate the need for specialized equipment like microarray printers.

Main Methods:

• A two-part microfluidic system was designed using polydimethylsiloxane (PDMS).
• A PDMS-based spotting device was used to array small interfering RNA (siRNA) molecules into 96 microwells.
• The siRNA array was transferred to a PDMS screening platform with microchannels for cell loading and transfection.

Main Results:

• The system successfully minimized reagent usage tenfold compared to conventional screening methods.
• The platform enabled efficient cell loading and transfection while preventing cross-contamination.
• Validation using Vesicular stomatitis virus and Rift Valley fever virus identified clathrin-mediated and caveolae-mediated endocytosis pathways, respectively.

Conclusions:

• The developed microfluidic system offers a cost-effective and miniaturized approach to RNAi screening.
• This platform facilitates the study of cellular pathways in virus entry and infection.
• The technique is adaptable for large-scale screening in various biosafety levels.