A novel and universal method for microRNA RT-qPCR data normalization
Pieter Mestdagh,Pieter Van Vlierberghe,An-Sofie De Weer,Daniel Muth,Frank Westermann,Frank Speleman,Jo Vandesompele +6 more
TLDR
It is demonstrated that the mean expression value outperforms the current normalization strategy in terms of better reduction of technical variation and more accurate appreciation of biological changes.Abstract:
Gene expression analysis of microRNA molecules is becoming increasingly important. In this study we assess the use of the mean expression value of all expressed microRNAs in a given sample as a normalization factor for microRNA real-time quantitative PCR data and compare its performance to the currently adopted approach. We demonstrate that the mean expression value outperforms the current normalization strategy in terms of better reduction of technical variation and more accurate appreciation of biological changes.read more
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Cell-free nucleic acids as biomarkers in cancer patients
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MicroRNA profiling: approaches and considerations.
TL;DR: This work reviews the major considerations for carrying out and interpreting results of miRNA-profiling studies and suggests several approaches that can be considered for effective use.
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miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis
Li Ma,Jennifer J. Young,Harsha K. Prabhala,Elizabeth Pan,Pieter Mestdagh,Daniel Muth,Julie Teruya-Feldstein,Ferenc Reinhardt,Tamer T. Onder,Scott Valastyan,Frank Westermann,Frank Speleman,Jo Vandesompele,Robert A. Weinberg +13 more
TL;DR: It is shown that miR-9, which is upregulated in breast cancer cells, directly targets CDH1, the E-cadherin-encoding messenger RNA, leading to increased cell motility and invasiveness, and a regulatory and signalling pathway involving a metastasis-promoting miRNA that is predicted to directly target expression of the key metastasis
miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis
Li Ma,Jennifer J. Young,Harsha K. Prabhala,Elizabeth Pan,Pieter Mestdagh,Daniel Muth,Julie Teruya-Feldstein,Ferenc Reinhardt,Tamer T. Onder,Scott Valastyan,Frank Westermann,Frank Speleman,Jo Vandesompele,Robert A. Weinberg +13 more
TL;DR: In this article, the authors uncover a regulatory and signalling pathway involving a metastasis-promoting miRNA that is predicted to directly target expression of the key metastasissuppressing protein E-cadherin.
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Adipose-derived circulating miRNAs regulate gene expression in other tissues
Thomas Thomou,Marcelo A. Mori,Jonathan M. Dreyfuss,Jonathan M. Dreyfuss,Masahiro Konishi,Masaji Sakaguchi,Christian Wolfrum,Tata Nageswara Rao,Tata Nageswara Rao,Jonathon N. Winnay,Ruben Garcia-Martin,Steven K. Grinspoon,Phillip Gorden,C. Ronald Kahn +13 more
TL;DR: Transplantation of both white and brown adipose tissue—brown especially—into ADicerKO mice restores the level of numerous circulating miRNAs that are associated with an improvement in glucose tolerance and a reduction in hepatic Fgf21 mRNA and circulating FGF21.
References
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Journal ArticleDOI
Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes
Jo Vandesompele,Katleen De Preter,Filip Pattyn,Bruce Poppe,Nadine Van Roy,Anne De Paepe,Franki Speleman +6 more
TL;DR: The normalization strategy presented here is a prerequisite for accurate RT-PCR expression profiling, which opens up the possibility of studying the biological relevance of small expression differences.
Journal ArticleDOI
The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments
Stephen A. Bustin,Vladimir Benes,Jeremy A. Garson,Jan Hellemans,Jim F. Huggett,Mikael Kubista,Reinhold Mueller,Tania Nolan,Michael W. Pfaffl,Gregory L. Shipley,Jo Vandesompele,Carl T. Wittwer,Carl T. Wittwer +12 more
TL;DR: The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines target the reliability of results to help ensure the integrity of the scientific literature, promote consistency between laboratories, and increase experimental transparency.
Journal ArticleDOI
MicroRNA expression profiles classify human cancers
Jun Lu,Gad Getz,Eric A. Miska,Eric A. Miska,Ezequiel Alvarez-Saavedra,Justin Lamb,David Peck,Alejandro Sweet-Cordero,Alejandro Sweet-Cordero,Benjamin L. Ebert,Benjamin L. Ebert,Raymond H. Mak,Raymond H. Mak,Adolfo A. Ferrando,James R. Downing,Tyler Jacks,H. Robert Horvitz,H. Robert Horvitz,Todd R. Golub,Todd R. Golub,Todd R. Golub +20 more
TL;DR: A new, bead-based flow cytometric miRNA expression profiling method is used to present a systematic expression analysis of 217 mammalian miRNAs from 334 samples, including multiple human cancers, and finds the miRNA profiles are surprisingly informative, reflecting the developmental lineage and differentiation state of the tumours.
Journal Article
Oncomirs : microRNAs with a role in cancer
TL;DR: I MicroRNAs (miRNAs) are an abundant class of small non-protein-coding RNAs that function as negative gene regulators as discussed by the authors, and have been shown to repress the expression of important cancer-related genes and might prove useful in the diagnosis and treatment of cancer.
Journal ArticleDOI
Normalization of Real-Time Quantitative Reverse Transcription-PCR Data: A Model-Based Variance Estimation Approach to Identify Genes Suited for Normalization, Applied to Bladder and Colon Cancer Data Sets
TL;DR: A novel, innovative, and robust strategy to identify stably expressed genes among a set of candidate normalization genes, rooted in a mathematical model of gene expression, that provides a direct measure for the estimated expression variation, enabling the user to evaluate the systematic error introduced when using the gene.
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