Quantitative analysis of miRNA expression in epithelial cells and tissues Journal Article
| Authors: | Bitzer, M.; Ju, W.; Jing, X.; Zavadil, J. |
| Article Title: | Quantitative analysis of miRNA expression in epithelial cells and tissues |
| Abstract: | Reliable detection of the microRNA (miRNA) precursor and mature form expression levels is a fundamental starting block for more focused studies of the biogenesis and functional roles of these important post-transcriptional modulators of gene expression. Building on our expertise with miRNA expression programs downstream of TGF-β/Smad signaling in homeostasis as well as in pathological conditions associated with epithelial tissues, we present a series of detailed and broadly applicable protocols for expression profiling of the mature miRNA forms using quantitative real-time PCR TaqMan, both single assays or low-density arrays. We next highlight key steps necessary for the detection of primary precursors of miRNAs (-pri-miRNAs) to address the initial steps of miRNA biogenesis, and we finally review some most widely used computational algorithms for miRNA target prediction used to complement experimental identification of the target mRNAs and proteins. © 2012 Springer Science+Business Media, LLC. |
| Keywords: | genetics; methodology; animal; metabolism; animals; microrna; gene expression; gene expression profiling; transforming growth factor beta; algorithms; rna; chemistry; algorithm; oligonucleotide array sequence analysis; epithelium cell; epithelial cells; real time polymerase chain reaction; dna microarray; micrornas; computer program; software; isolation and purification; epithelial tissue; epithelial cell; mirna; mirna expression; real-time polymerase chain reaction; mirna target prediction; pri-mirna expression; primer3; quantitative real-time pcr; sybr green pcr; taqman array microrna card; taqman microrna assay; tgf-? |
| Journal Title: | Methods in Molecular Biology |
| Volume: | 820 |
| ISSN: | 1064-3745 |
| Publisher: | Humana Press Inc |
| Date Published: | 2012-01-01 |
| Start Page: | 55 |
| End Page: | 70 |
| Language: | English |
| DOI: | 10.1007/978-1-61779-439-1_4 |
| PROVIDER: | scopus |
| PUBMED: | 22131025 |
| DOI/URL: | |
| Notes: | Chapter 4 in "Cytokine Protocols" (ISBN: 978-1-61779-438-4) --- Export Date: 1 February 2012 - Source: Scopus |
