Induced nitric oxide synthase (iNOS) is an enzyme that catalyzes the production of nitric oxide in many eukaryotic species. iNOS expression is up-regulated in inflammation, making it a good biomarker for monitoring injured tissue. We have investigated the use of strand-displacement activated fluorescent probes to image and quantitatively report the expression level of iNOS mRNA in vitro. The probes are constructed by annealing an antisense PNA bearing fluorescein to a shorter DNA sequence bearing Dabcyl which quenches the fluorescence of the fluorescein.  In thepresence of the target mRNA, however, the mRNA will bind to the complementaryPNA and displace the shorter DNA sequence, thereby restoring fluorescence ofthe fluorescein. The fluorescence of matched probe increases almost 10 times in the presence of a complementary DNA. We will report on the ability of the probes to detectthe expression of iNOS mRNA sequence in both fixed and live RAW 264.7cells.

Strand-displacement activated fluorescent
PNA probes for detecting iNOS mRNA expression

240th National Meeting (2010)

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