Profiling microRNA expression using sensitive cDNA probes and filter arrays.
MicroRNAs (miRNAs) are small noncoding RNAs (approximately 22 nucleotides) that have recently emerged as important regulators of gene expression in both plants and animals. With few exceptions, however, the target genes and the expression levels of most miRNAs are unknown. Here we show that direct random-primed cDNA synthesis on either chemically synthesized small RNAs (21-22 nucleotides) or gel-purified mature miRNAs from human cells can produce specific and sensitive full-length cDNA probes. Using oligonucleotide filter arrays, we demonstrate that the internally labeled cDNA probes are sensitive for detecting differential miRNA expression between untreated and O-tetradecanoylphorbol-13-acetate (TPA)-treated HL60 cells. The present study should facilitate a high-throughput analysis of miRNA expression between samples.
Sioud
Mouldy
M
Department of Immunology, Institute for Cancer Research, The Norwegian Radium Hospital, Oslo, Norway. mosioud@ulrik.uio.no
Røsok
Oystein
O
D015342:DNA Probes / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D066298:In Vitro Techniques; D035683:MicroRNAs / Q000032:analysis*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*
D015342:DNA Probes; D018076:DNA, Complementary; D035683:MicroRNAs