Imaging target mRNA and siRNA-mediated gene silencing in vivo with ribozyme-based reporters. Export

@article{citeulike:4371091, abstract = {Noninvasive imaging of specific mRNAs in living subjects promises numerous biological and medical applications. Common strategies use fluorescently or radioactively labelled antisense probes to detect target mRNAs through a hybridization mechanism, but have met with limited success in living animals. Here we present a novel molecular imaging approach based on the group I intron of Tetrahymena thermophila for imaging mRNA molecules in vivo. Engineered trans-splicing ribozyme reporters contain three domains, each of which is designed for targeting, splicing, and reporting. They can transduce the target mRNA into a reporter mRNA, leading to the production of reporter enzymes that can be noninvasively imaged in vivo. We have demonstrated this ribozyme-mediated RNA imaging method for imaging a mutant p53 mRNA both in single cells and noninvasively in living mice. After optimization, the ribozyme reporter increases contrast for the transiently expressed target by 180-fold, and by ten-fold for the stably expressed target. siRNA-mediated specific gene silencing of p53 expression has been successfully imaged in real time in vivo. This new ribozyme-based RNA reporter system should open up new avenues for in vivo RNA imaging and direct imaging of siRNA inhibition.}, author = {So, M. K. and Gowrishankar, G. and Hasegawa, S. and Chung, J. K. and Rao, J.}, citeulike-article-id = {4371091}, citeulike-linkout-0 = {http://dx.doi.org/10.1002/cbic.200800370}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/18972511}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=18972511}, citeulike-linkout-3 = {http://www3.interscience.wiley.com/cgi-bin/abstract/121493694/ABSTRACT}, day = {3}, doi = {10.1002/cbic.200800370}, issn = {1439-7633}, journal = {Chembiochem : a European journal of chemical biology}, keywords = {detection, reporter, ribozyme, rna}, month = {November}, number = {16}, pages = {2682--2691}, posted-at = {2009-04-21 03:55:50}, priority = {0}, title = {Imaging target mRNA and siRNA-mediated gene silencing in vivo with ribozyme-based reporters.}, url = {http://dx.doi.org/10.1002/cbic.200800370}, volume = {9}, year = {2008} }

TY - JOUR ID - citeulike:4371091 L3 - citeulike-article-id:4371091 N2 - Noninvasive imaging of specific mRNAs in living subjects promises numerous biological and medical applications. Common strategies use fluorescently or radioactively labelled antisense probes to detect target mRNAs through a hybridization mechanism, but have met with limited success in living animals. Here we present a novel molecular imaging approach based on the group I intron of Tetrahymena thermophila for imaging mRNA molecules in vivo. Engineered trans-splicing ribozyme reporters contain three domains, each of which is designed for targeting, splicing, and reporting. They can transduce the target mRNA into a reporter mRNA, leading to the production of reporter enzymes that can be noninvasively imaged in vivo. We have demonstrated this ribozyme-mediated RNA imaging method for imaging a mutant p53 mRNA both in single cells and noninvasively in living mice. After optimization, the ribozyme reporter increases contrast for the transiently expressed target by 180-fold, and by ten-fold for the stably expressed target. siRNA-mediated specific gene silencing of p53 expression has been successfully imaged in real time in vivo. This new ribozyme-based RNA reporter system should open up new avenues for in vivo RNA imaging and direct imaging of siRNA inhibition. IS - 16 JF - Chembiochem : a European journal of chemical biology SN - 1439-7633 EP - 2691 TI - Imaging target mRNA and siRNA-mediated gene silencing in vivo with ribozyme-based reporters. VL - 9 SP - 2682 KW - detection KW - reporter KW - ribozyme KW - rna AU - So, MK AU - Gowrishankar, G AU - Hasegawa, S AU - Chung, JK AU - Rao, J PY - 2008/11/03/ UR - http://dx.doi.org/10.1002/cbic.200800370 ER -