Design, preparation and testing of suitable probe-receptors for RNA biosensing

Area of Science:

• Biotechnology
• Molecular Biology
• Biosensor Technology

Background:

• Conventional molecular biology methods for RNA quantification present operative and analytical limitations.
• There is a need for cheaper, easier, rapid, and reproducible methods for absolute RNA measurements.

Purpose of the Study:

• To develop an integrated approach for designing, synthesizing, and connecting RNA probes to a microgravimetric biosensor.
• To create a biosensor capable of overcoming the limitations of existing RNA measurement techniques.

Main Methods:

• A computational method was used to design RNA probes based on target accessibility, duplex stability, and sequence uniqueness.
• Automated chemical synthesis produced selected oligonucleotide probes.
• Probes were covalently conjugated to the sensing surface of a quartz microbalance.
• The microgravimetric biosensor was tested using synthetic and full-length mRNA targets in a flow chamber.

Main Results:

• Specific, dose-dependent binding of synthetic target substrates was observed.
• The successful monitoring of full-length mRNA binding demonstrated the biosensor's capability.
• The developed biosensor technology enables absolute RNA measurements.

Conclusions:

• The integrated RNA biosensor design, synthesis, and conjugation approach is effective.
• This biosensor technology offers a viable solution for overcoming conventional method limitations.
• The developed biosensor provides a cheap, easy, rapid, and reproducible method for absolute RNA quantification.