A biochemical approach to identifying microRNA targets Export

@article{citeulike:2023141, abstract = {10.1073/pnas.0709971104 Identifying the downstream targets of microRNAs (miRNAs) is essential to understanding cellular regulatory networks. We devised a direct biochemical method for miRNA target discovery that combined RNA-induced silencing complex (RISC) purification with microarray analysis of bound mRNAs. Because targets of miR-124a have been analyzed, we chose it as our model. We honed our approach both by examining the determinants of stable binding between RISC and synthetic target RNAs and by determining the dependency of both repression and RISC coimmunoprecipitation on miR-124a seed sites in two of its well characterized targets . Examining the complete spectrum of miR-124 targets in 293 cells yielded both a set that were down-regulated at the mRNA level, as previously observed, and a set whose mRNA levels were unaffected by miR-124a. Reporter assays validated both classes, extending the spectrum of mRNA targets that can be experimentally linked to the miRNA pathway.}, author = {Karginov, Fedor V. and Conaco, Cecilia and Xuan, Zhenyu and Schmidt, Bryan H. and Parker, Joel S. and Mandel, Gail and Hannon, Gregory J.}, citeulike-article-id = {2023141}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0709971104}, citeulike-linkout-1 = {http://www.pnas.org/content/104/49/19291.abstract}, citeulike-linkout-2 = {http://www.pnas.org/content/104/49/19291.full.pdf}, citeulike-linkout-3 = {http://www.pnas.org/cgi/content/abstract/104/49/19291}, citeulike-linkout-4 = {http://view.ncbi.nlm.nih.gov/pubmed/18042700}, citeulike-linkout-5 = {http://www.hubmed.org/display.cgi?uids=18042700}, day = {4}, doi = {10.1073/pnas.0709971104}, journal = {Proceedings of the National Academy of Sciences}, keywords = {biochemical\_method, microrna, target}, month = {December}, number = {49}, pages = {19291--19296}, posted-at = {2009-01-12 02:34:27}, priority = {2}, title = {A biochemical approach to identifying microRNA targets}, url = {http://dx.doi.org/10.1073/pnas.0709971104}, volume = {104}, year = {2007} }

TY - JOUR ID - citeulike:2023141 L3 - citeulike-article-id:2023141 N2 - 10.1073/pnas.0709971104 Identifying the downstream targets of microRNAs (miRNAs) is essential to understanding cellular regulatory networks. We devised a direct biochemical method for miRNA target discovery that combined RNA-induced silencing complex (RISC) purification with microarray analysis of bound mRNAs. Because targets of miR-124a have been analyzed, we chose it as our model. We honed our approach both by examining the determinants of stable binding between RISC and synthetic target RNAs and by determining the dependency of both repression and RISC coimmunoprecipitation on miR-124a seed sites in two of its well characterized targets . Examining the complete spectrum of miR-124 targets in 293 cells yielded both a set that were down-regulated at the mRNA level, as previously observed, and a set whose mRNA levels were unaffected by miR-124a. Reporter assays validated both classes, extending the spectrum of mRNA targets that can be experimentally linked to the miRNA pathway. IS - 49 JF - Proceedings of the National Academy of Sciences EP - 19296 TI - A biochemical approach to identifying microRNA targets VL - 104 SP - 19291 KW - biochemical_method KW - microrna KW - target AU - Karginov, Fedor AU - Conaco, Cecilia AU - Xuan, Zhenyu AU - Schmidt, Bryan AU - Parker, Joel AU - Mandel, Gail AU - Hannon, Gregory PY - 2007/12/04/ UR - http://dx.doi.org/10.1073/pnas.0709971104 ER -