A Real-time Biosensor of Antiproliferative Immune Messenger 2-5A for Research, Diagnostics, and High-throughput Screening

Web Published:

Princeton Docket # 17-3323-1


Princeton researchers have engineered a light emitting reporter to measure the real-time buildup of 2’-5’ linked oligoadenylates (2-5As) biochemically and in live cells using a one-step, high-throughput method.

2-5As are produced in stressed human cells that accumulate double stranded RNA (dsRNA). 2-5A serves as specific activators of cellular RNA processing by kinase-ribonuclease RNase L, which rapidly stops cell-wide translation. 2-5A levels can serve as a marker of dsRNA presence, i.e. to assess viral infections and cell stresses associated with altered migration, metastasis and cancers. The light emitting reporter allows one to continuously observe 2-5A production in cells exposed to poly-IC, a dsRNA mimetic. This tool is directly suitable for high-throughput screens for small molecule modulators of the 2-5A/RNase L axis. Unlike existing technologies, this reporter can be used in live animals for 2-5A detection.



•       Disease monitoring and diagnostic tool

•       Treatment success monitoring

•       Screening for small molecule modulators of 2-5/RNase L system

•       In vivo studies in cells and animals



•       Real-time  continuous 2-5A monitoring

•       Single-protein design

•       Uses basic equipment

•       Scalable for high-throughput use

•       Compatible with cells

•       The only non-invasive 2-5A biosensor


Stage of Development:


The reporter has been shown to respond to 2-5A in proof-of-concept biochemical assays and in genetically altered Hela cells and in A549 human lung cancer cells as verified by western blot.


The Inventors:


Alexei Korennykh, Ph.D. is an Associate Professor of Molecular Biology. The central theme of Prof. Korennykh’s research is to understand mechanisms of RNA-dependent signal transduction in the human immune system, and during stress responses associated with human diseases. His lab uses X-ray crystallography, biochemistry, biophysics, and cell biology to understand structures, functions and regulation of the key human proteins, RNA, intracellular messengers, and their complexes.


Alisha Chitrakar is a Ph.D. student in the Korennykh Lab. Alisha is interested in understanding mechanisms of stress responses in the human innate immune system and second messengers that relay signals of immunogenic stress. She has expertise in RNA biology and cell biology.


Jesse Donovan, Ph.D., is an associate research scholar in the Korennykh lab.  Dr. Donovan is broadly interested in RNA biology with a focus on the post-transcriptional control of gene expression.  Highlights of his work in the Korennykh group include uncovering the structural basis for the activation of the 2-5A synthesizing enzymes and the development of a high-throughput sequencing strategy to discover RNAs cleaved by RNase L and other metal-independent RNases.


Intellectual Property Status:

Patent protection is pending.

Princeton is seeking to identify appropriate partners for the further development and commercialization of this technology.




Linda Jan

Princeton University Office of Technology Licensing • (609) 258-5579 • ljan@princeton.edu


Anthony Williams

Princeton University Office of Technology Licensing • (609) 258-3769 • anthonyw@princeton.edu



Patent Information:
For Information, Contact:
Prabhpreet Gill
Licensing Associate
Princeton University
Alexei Korennykh
Alisha Chitrakar
Jesse Donovan