Identification Of Single Nucleotide Polymorphisms In MicroRNA 146A

Abstract

MicroRNAs and other small noncoding RNAs regulate nearly 50% of genes at either the transcriptional or Rationale and Objectives: post-transcriptional levels. It is therefore not surprising that the expression of the miRNA genes themselves is tightly regulated. We previously implicated differential expression of miRNA 146a in COPD and control fibroblasts and showed that its target site in the 3'-UTR of COX-2 regulates levels of this protein, and consequent PGE production. We hypothesize that differential expression of miRNA 146a in COPD and control fibroblasts may be due to single nucleotide polymorphisms in this gene.Genomic DNA was isolated from 10 fibroblast strains obtained from the human lungs of control and COPD subjects. Material and Methods: The complete coding region of the pre-miRNA146a (99bp) as well as 5 kb of 5' genomic and 1 kb of 3'genomic was resequenced.A total of 3 single nucleotide polymorphisms were identified, all of which are novel. The SNPs were located at -853A/C, -1135G/A Results: and -3014G/A relative to the pre-miRNA 146a start site. Within 25 bp of the -1135 and -3014 polymorphic sites, the OCT1 transcription factor was predicted to have multiple binding sites (MatInspector prediction software 8.0). The polymorphism at -853 had no predicted transcription factor binding sites.These data suggest that single nucleotide polymorphisms are common in the miRNA146a regulatory region. In the future we Conclusion: intend to screen a total of 20 control and 20 COPD fibroblast strains for single nucleotide polymorphisms and relate these polymorphisms to the clinical phenotypes of COPD. This abstract is funded by: State of Nebraska LB506 Am J Respir Crit Care Med 183;2011:A3526 Internet address: www.atsjournals.org Online Abstracts Issue