title: MicroRNA target prediction: theory and practice.
abstract: The present study is one of the few that includes tissue samples in the evaluation of target prediction algorithms designed to detect microRNA (miRNA) sequences that might interact with particular messenger RNA (mRNA) sequences. Twelve different target prediction tools were used to find miRNA sequences that might interact with CCL20 gene expression. Different algorithms predicted controversial miRNA sequences for CCL20 regulation due to a different weighting of parameters. Hsa-miR-21 and hsa-miR-145 suggested by four or more programs were chosen for further investigation. Possible real interaction of these miRNA sequences with CCL20 gene expression was monitored using luciferase assays and expression analyses of tissue samples of colorectal adenocarcinoma by either qRT-PCR or ELISA. Folding status of seed-binding sites in complete mRNA and 3'UTR of CCL20 was predicted. Prediction of miRNA expression was attempted based on CCL20 expression data. Eight of the target prediction tools forecasted a role for hsa-miR-21 and four mentioned hsa-miR-145 in CCL20 gene regulation. Laboratory experimentation showed that CCL20 may serve as a target of hsa-miR-21 but not hsa-miR-145. Expression of the molecules resulted in no clear assertion. Folding of seed-binding sites was predicted to be relatively constant for the complete mRNA and 3'UTR. Predicting miRNA expression based on target gene expression was impossible. This might be attributable to the fact that effects of miRNA activity may oscillate between gene product repression and activation. Additional systematic studies are needed to address this issue.
authors.affiliation: Department of Pathology, University of Saarland Medical School, Homburg Saar, Germany.
authors.forename: Mathias
authors.initials: M
authors.lastname: Wagner
authors.forename: Benjamin
authors.initials: B
authors.lastname: Vicinus
authors.forename: Vilma Oliveira
authors.initials: VO
authors.lastname: Frick
authors.forename: Michael
authors.initials: M
authors.lastname: Auchtor
authors.forename: Claudia
authors.initials: C
authors.lastname: Rubie
authors.forename: Pascal
authors.initials: P
authors.lastname: Jeanmonod
authors.forename: Tereza A
authors.initials: TA
authors.lastname: Richards
authors.forename: Roland
authors.initials: R
authors.lastname: Linder
authors.forename: Frank
authors.initials: F
authors.lastname: Weichert
mesh_terms: D020413:3' Untranslated Regions; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000465:Algorithms; D001665:Binding Sites; D054418:Chemokine CCL20 / Q000235:genetics*; D015179:Colorectal Neoplasms / Q000235:genetics / Q000401:mortality; D005260:Female; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000737:chemistry / Q000378:metabolism*; D008875:Middle Aged; D059370:RNA Folding; D012333:RNA, Messenger / Q000737:chemistry* / Q000378:metabolism; D038621:Regulatory Sequences, Ribonucleic Acid; D012984:Software
chemical_list: D020413:3' Untranslated Regions; C108355:CCL20 protein, human; D054418:Chemokine CCL20; C541010:MIRN145 microRNA, human; C524940:MIRN21 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D038621:Regulatory Sequences, Ribonucleic Acid