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. 2004 Jun 29;101(26):9740-4.
doi: 10.1073/pnas.0403293101. Epub 2004 Jun 21.

An oligonucleotide microchip for genome-wide microRNA profiling in human and mouse tissues

Chang-Gong Liu  1 , George Adrian Calin, Brian Meloon, Nir Gamliel, Cinzia Sevignani, Manuela Ferracin, Calin Dan Dumitru, Masayoshi Shimizu, Simona Zupo, Mariella Dono, Hansjuerg Alder, Florencia Bullrich, Massimo Negrini, Carlo M Croce
Affiliations

An oligonucleotide microchip for genome-wide microRNA profiling in human and mouse tissues

Chang-Gong Liu et al. Proc Natl Acad Sci U S A. .

Abstract

MicroRNAs (miRNAs) are a class of small noncoding RNA genes recently found to be abnormally expressed in several types of cancer. Here, we describe a recently developed methodology for miRNA gene expression profiling based on the development of a microchip containing oligonucleotides corresponding to 245 miRNAs from human and mouse genomes. We used these microarrays to obtain highly reproducible results that revealed tissue-specific miRNA expression signatures, data that were confirmed by assessment of expression by Northern blots, real-time RT-PCR, and literature search. The microchip oligolibrary can be expanded to include an increasing number of miRNAs discovered in various species and is useful for the analysis of normal and disease states.

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Figures

Fig. 1.
Fig. 1.
Microarray expression data. (a) miRNA expression in HeLa cells by using various quantities of total RNA. (b) miRNA expression in human and mouse samples: human peripheral blood leucocytes (PBL) from three healthy donors and two samples of mouse RAW 264.7 macrophages. Five micrograms of total RNA were used and hybridized as described.
Fig. 2.
Fig. 2.
The human miRNome expression profiles corresponding to 161 miRNAs. (a) Distinct patterns of miRNA expression found in human adult and fetal tissues. (b) Examples of miRNA signatures: specific overexpression of various miRNAs in skeletal muscle, heart, prostate, and brain. All of the data represent the average of at least two replicates or two samples from the same tissue.
Fig. 3.
Fig. 3.
Northern blot confirmation of the microarray data. The names of miRNAs and specific oligonucleotides spotted on the array are presented. The numbers correspond to absolute expression value of each miRNA (determined by a per-chip on median normalization) and, therefore, can be compared with the band intensity on Northern blots. 5S rRNA stained with ethidium bromide was used as loading control.
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