Type

Journal Article

Authors

Catherine M Greene
Noel G McElvaney
Shane O'Neill
Irene K Oglesby
Kevin Gaughan
Stephen Smith
Tidi Hassan

Subjects

Microbiology

Topics
rna medicine and health sciences messenger interleukin 8 mrna expression alpha 1 antitrypsin targeting sequences rna precursors rna messenger cell lines secretory leukocyte peptidase inhibitor isolation purification transcriptome binding sites prediction models reverse transcriptase polymerase chain reaction cell line cell humans antisense oligodeoxyribonucleotides antisense micrornas immunology inflammation infection biotinylation oligodeoxyribonucleotides genetics enzyme linked immunosorbent assay metabolism rna isolation

Isolation and identification of cell-specific microRNAs targeting a messenger RNA using a biotinylated anti-sense oligonucleotide capture affinity technique. (2013)

Abstract MicroRNAs (miRNAs) are small non-coding RNAs that regulate expression by translational repression or messenger RNA (mRNA) degradation. Although numerous bioinformatic prediction models exist to identify miRNA-mRNA interactions, experimental validation of bona fide interactions can be difficult and laborious. Few methods can comprehensively identify miRNAs that target a single mRNA. We have developed an experimental approach to search for miRNAs targeting any mRNA using a capture affinity assay involving a biotinylated DNA anti-sense oligonucleotide. This method identifies miRNAs targeting the full length of the mRNA. The method was tested using three separate mRNA targets: alpha-1 antitrypsin (AAT) mRNA, interleukin-8 mRNA and secretory leucoprotease inhibitor mRNA. AAT mRNA-specific and total miRNAs from three different cell lines (monocytic THP-1, bronchial epithelial 16HBE14o- and liver HepG2 cells) were profiled, and validation studies revealed that AAT mRNA-specific miRNAs functionally target the AAT mRNA in a cell-specific manner, providing the first evidence of innate miRNAs selectively targeting and modulating AAT mRNA expression. Interleukin-8 and secretory leucoprotease inhibitor mRNAs and their cognate miRNAs were also successfully captured using this approach. This is a simple and an efficient method to potentially identify miRNAs targeting sequences within the full length of a given mRNA transcript.
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Full list of authors on original publication

Catherine M Greene, Noel G McElvaney, Shane O'Neill, Irene K Oglesby, Kevin Gaughan, Stephen Smith, Tidi Hassan

Experts in our system

1
Catherine M Greene
Royal College of Surgeons in Ireland
Total Publications: 150
 
2
Noel G McElvaney
Royal College of Surgeons in Ireland
Total Publications: 194
 
3
Shane J O'Neill
Royal College of Surgeons in Ireland
Total Publications: 84
 
4
Stephen Smith
Trinity College Dublin
Total Publications: 27
 
5
Tidi Hassan
Royal College of Surgeons in Ireland