title: Housekeeping gene selection advisory: glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin are targets of miR-644a.
abstract: 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 β-actin have been used extensively for normalization of gene expression data. Here, we show that GAPDH and β-actin are direct targets of miR-644a. Our data demonstrate the unsuitability of GAPDH and β-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.
authors.affiliation: Department of Biological, Geological and Environmental Sciences, Cleveland State University, Cleveland, Ohio, United States of America.
authors.forename: Kavleen
authors.initials: K
authors.lastname: Sikand
authors.forename: Jagjit
authors.initials: J
authors.lastname: Singh
authors.forename: Jey Sabith
authors.initials: JS
authors.lastname: Ebron
authors.forename: Girish C
authors.initials: GC
authors.lastname: Shukla
mesh_terms: D020413:3' Untranslated Regions; D000199:Actins / Q000235:genetics*; D020029:Base Pairing; D001483:Base Sequence; D002460:Cell Line; D005786:Gene Expression Regulation; D023061:Gene Order; D020043:Genes, Essential; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D020413:3' Untranslated Regions; D000199:Actins; C579242:MIRN644 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases