Housekeeping Gene Selection Advisory: Glyceraldehyde-3-Phosphate Dehydrogenase and [beta]-Actin Are Targets of miR-644a

Housekeeping Gene Selection Advisory: Glyceraldehyde-3-Phosphate Dehydrogenase and [beta]-Actin Are Targets of miR-644a

Results of overexpression or downregulation of a microRNA (miRNA) on its target mRNA expression are often validated by reverse-transcription and quantitative PCR analysis using an appropriate housekeeping gene as an internal control. The possible direct or indirect effects of a miRNA on the expressi...

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Veröffentlicht in:PloS one 2012-10, Vol.7 (10), p.e47510
Hauptverfasser: Sikand, Kavleen, Singh, Jagjit, Ebron, Jey Sabith, Shukla, Girish C
Format: Artikel
Sprache:eng
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Actin
Chemical properties
Gene expression
Glyceraldehyde 3-phosphate dehydrogenase
MicroRNA
Polymerase chain reaction
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creator Sikand, Kavleen
Singh, Jagjit
Ebron, Jey Sabith
Shukla, Girish C
description Results of overexpression or downregulation of a microRNA (miRNA) on its target mRNA expression are often validated by reverse-transcription and quantitative PCR analysis using an appropriate housekeeping gene as an internal control. The possible direct or indirect effects of a miRNA on the expression of housekeeping genes are often overlooked. Among many housekeeping genes, expressions of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and [beta]-actin have been used extensively for normalization of gene expression data. Here, we show that GAPDH and [beta]-actin are direct targets of miR-644a. Our data demonstrate the unsuitability of GAPDH and [beta]-actin as internal controls in miR-644a functional studies and emphasize the need to carefully consider the choice of a reference gene in miRNA experiments.
doi_str_mv 10.1371/journal.pone.0047510
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subjects Actin
Chemical properties
Gene expression
Glyceraldehyde 3-phosphate dehydrogenase
MicroRNA
Polymerase chain reaction
title Housekeeping Gene Selection Advisory: Glyceraldehyde-3-Phosphate Dehydrogenase and [beta]-Actin Are Targets of miR-644a
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