Dal trascrittoma all’interattoma di miRNA: identificazione sperimentale e bioinformatica delle interazioni funzionali miRNA:mRNA

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Abstract

[automatically translated] MiRNAs, small endogenous non-coding RNA molecules, regulate gene expression through the degradation of the messenger (mRNA) or inhibition of translation. The mature miRNAs interact with the proteins of the RISC complex (RNA-induced silencing complex) including the Argonaute protein (Aug), capable of binding directly miRNAs and to mediate the regulation of gene expression following the interaction of miRNAs with its own target mRNA. A single miRNA can bind different mRNA and each mRNA can be regulated by several miRNAs. Most of prediction software available today identify the putative targets of individual miRNAs ignoring global type of characteristics, such as the levels of mRNA expression, and miRNAs coexpressed. The aim of our work is to develop a system for the study and prediction of miRNA functional interactions: mRNA that takes account of all miRNA and mRNA coexpressed in a cell under certain conditions. In order to measure and relate the levels of mRNA and miRNA, we have developed a highly efficient immunoprecipitation of the complex RISC-Ago2 from cytoplasmic extracts of breast cancer cell line MCF-7. The profiles of expression of mRNA and miRNA samples obtained were obtained by microarray. This technique allowed us to separately characterize the set of mRNA and miRNA bound and unbound to the RISC complex. And 'centrifugation on density gradient was also used to separate the mRNA being translated from those not translated and miRNAs associated with each of these fractions. Even these samples will be analyzed by microarray. The analysis of the data obtained enable us to identify, among all miRNAs and mRNAs expressed in the cell those involved in the adjustment process and identify the interactions miRNA: mRNA that induce a translation of the block or the degradation of the target messenger. Using the data obtained we propose to develop a mathematical model to predict the set of functional interactions between the miRNA and its target cell in any condition in which they are known the expression profiles of miRNAs and mRNAs. Even these samples will be analyzed by microarray. The analysis of the data obtained enable us to identify, among all miRNAs and mRNAs expressed in the cell those involved in the adjustment process and identify the interactions miRNA: mRNA that induce a translation of the block or the degradation of the target messenger. Using the data obtained we propose to develop a mathematical model to predict the set of functional interactions between the miRNA and its target cell in any condition in which they are known the expression profiles of miRNAs and mRNAs. Even these samples will be analyzed by microarray. The analysis of the data obtained enable us to identify, among all miRNAs and mRNAs expressed in the cell those involved in the adjustment process and identify the interactions miRNA: mRNA that induce a translation of the block or the degradation of the target messenger. Using the data obtained we propose to develop a mathematical model to predict the set of functional interactions between the miRNA and its target cell in any condition in which they are known the expression profiles of miRNAs and mRNAs. a translation of the mRNA which induce or block the degradation of the target messenger. Using the data obtained we propose to develop a mathematical model to predict the set of functional interactions between the miRNA and its target cell in any condition in which they are known the expression profiles of miRNAs and mRNAs. a translation of the mRNA which induce or block the degradati
Original languageItalian
Number of pages58
Publication statusPublished - 2013

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