title: Analysis of microRNA signatures using size-coded ligation-mediated PCR.
abstract: The expression pattern and regulatory functions of microRNAs (miRNAs) are intensively investigated in various tissues, cell types and disorders. Differential miRNA expression signatures have been revealed in healthy and unhealthy tissues using high-throughput profiling methods. For further analyses of miRNA signatures in biological samples, we describe here a simple and efficient method to detect multiple miRNAs simultaneously in total RNA. The size-coded ligation-mediated polymerase chain reaction (SL-PCR) method is based on size-coded DNA probe hybridization in solution, followed-by ligation, PCR amplification and gel fractionation. The new method shows quantitative and specific detection of miRNAs. We profiled miRNAs of the let-7 family in a number of organisms, tissues and cell types and the results correspond with their incidence in the genome and reported expression levels. Finally, SL-PCR detected let-7 expression changes in human embryonic stem cells as they differentiate to neuron and also in young and aged mice brain and bone marrow. We conclude that the method can efficiently reveal miRNA signatures in a range of biological samples.
authors.affiliation: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran 1585636473, Iran.
authors.forename: Ehsan
authors.initials: E
authors.lastname: Arefian
authors.forename: Jafar
authors.initials: J
authors.lastname: Kiani
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.forename: S Ali M
authors.initials: SA
authors.lastname: Shariati
authors.forename: Seyed Hamid
authors.initials: SH
authors.lastname: Aghaee-Bakhtiari
authors.forename: Amir
authors.initials: A
authors.lastname: Atashi
authors.forename: Yousof
authors.initials: Y
authors.lastname: Gheisari
authors.forename: Naser
authors.initials: N
authors.lastname: Ahmadbeigi
authors.forename: Ali M
authors.initials: AM
authors.lastname: Banaei-Moghaddam
authors.forename: Mahmood
authors.initials: M
authors.lastname: Naderi
authors.forename: Nabiolah
authors.initials: N
authors.lastname: Namvarasl
authors.forename: Liam
authors.initials: L
authors.lastname: Good
authors.forename: Omid R
authors.initials: OR
authors.lastname: Faridani
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001853:Bone Marrow / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D011088:DNA Ligases; D053595:Embryonic Stem Cells / Q000378:metabolism; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000032:analysis* / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D012322:RNA Precursors / Q000032:analysis
chemical_list: D015415:Biomarkers; D035683:MicroRNAs; D012322:RNA Precursors; C487931:mirnlet7 microRNA, human; D011088:DNA Ligases