clampFISH probes for high-grain, single molecule amplification of RNA

A non-enyzmatic, high-gain signal amplification method for fluorescent detection of RNA to achieve greater than 400 times signal amplification. This makes signal detection possible under low-magnification and separation of cells by RNA levels via flow cytometry.

Technology Overview:

Many amplification approaches rely on enzymes to achieve amplification. However, these for single nucleic acid amplification, existing approaches are non-specific and do not readily allow for measurements of large populations of cells for heterogeneity in gene expression. Alternative systems for single nucleic acid amplification require unique microscopy techniques.

Researchers at Penn have demonstrated, through the use of click chemistry, that their clampFISH can achieve exponential amplification of target probes that detect single nucleic acids. Over 6 rounds of amplification they have achieved a 120-fold increase for RNA with very low background noise. This process makes the amplification technique viable for high throughput methods (flow cytometer) for measurement of single transcripts within a cell. The technology is currently being translated to multiplexed applications.

Applications:

Amplification for high-throughput and possibly multiplexing systems

Advantages:

• High Amplification of Single Nucleic Acid in Tissue
• Low Background Noise
• Lends itself to high throughput and multiplexed applications
• Relieves the use of non-specific, bulky enzymes (less steps to achieve desired amplification)
• No need for high-powered camera

Stage of Development:

• Prototype and Demonstration Model Developed
• Multiplexed Demonstrated

Intellectual Property:

Patent Pending

Desired Partnerships:

• License
• Co-development (this replaces collaboration or sponsored research)

Docket # 17-8198