A Novel Approach for Designing in Situ Hybrization Probes

"FISH-STICs” offers an RNA-FISH probe synthesis approach that is low cost and easy accessible

In vitro–transcribed in situ hybridization (ISH) probes have long been applied in histology, and whole-mount gene expression pattern analysis, but they have found little application in the single-molecule RNA detection desirable in studies of mRNA localization. For that purpose, multiply labeled fluorescent Oligo Deoxy Nucleotides (ODN) probes can image single mRNAs (RNA-fluorescence in situ hybridization, RNA-FISH), however, the synthesis of FISH currently requires in house DNA synthesis and post-synthesis dye coupling, an inefficient process that is difficult to control and therefore inaccessible to most laboratories. Thus, alternative methods of making FISH more accessible and sensitive are highly sought-after.

Dr. Kevin Czaplinski has developed a strategy that uses commercially synthesized oligonucleotides as RNA-FISH probes without further modification and show that such probes work well for detection of RNA in cultured cells. His approach - namely FISH with Sequential Tethered and Intertwined nucleic acid molecule Complexes (FISH-STICs) - can bind a high concentration of fluorescent ODN to a short stretch of an RNA using commercial DNA synthesis outlets available to any laboratory. The situ hybridization probes FISH-STICs overcome the above noted limitations and permit rapid, simple, cost efficient and sensitive detection of multiple nucleic acids (and/or other nucleic acids) simultaneously.

One FISH-STIC probe detects mRNA molecules in culture, probe detection can be improved by the addition of multiple probes that can be easily adapted for robust mRNA quantification Rapid, simple and cost-effective

Research Tool Diagnostics

US-2015-0105298

Research

We seek to develop and commercialize, by an exclusive or non-exclusive license agreement with a company active in the area

Available for License

Sinnamon, J. R., & Czaplinski, K. (2014). rna, 20(2), 260-266.

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