Experimental Procedures to Identify and Validate Specific MRNA Targets of MiRNAs
Authors
Affiliations
Functionally matured microRNAs (miRNAs) are small single-stranded non-coding RNA molecules which are emerging as important post-transcriptional regulators of gene expression and consequently are central players in many physiological and pathological processes. Since the biological roles of individual miRNAs will be dictated by the mRNAs that they regulate, the identification and validation of miRNA/mRNA target interactions is critical for our understanding of the regulatory networks governing biological processes. We promulgate the combined use of prediction algorithms, the examination of curated databases of experimentally supported miRNA/mRNA interactions, manual sequence inspection of cataloged miRNA binding sites in specific target mRNAs, and review of the published literature as a reliable practice for identifying and prioritizing biologically important miRNA/mRNA target pairs. Once a preferred miRNA/mRNA target pair has been selected, we propose that the authenticity of a functional miRNA/mRNA target pair be validated by fulfilling four well-defined experimental criteria. This review summarizes our current knowledge of miRNA biology, miRNA/mRNA target prediction algorithms, validated miRNA/mRNA target data bases, and outlines several experimental methods by which miRNA/mRNA targets can be authenticated. In addition, a case study of human endoglin is presented as an example of the utilization of these methodologies.
In silico prioritisation of microRNA-associated common variants in multiple sclerosis.
Fashina I, McCoy C, Furney S Hum Genomics. 2023; 17(1):31.
PMID: 36991503 PMC: 10061723. DOI: 10.1186/s40246-023-00478-4.
Trinidad-Barnech J, Fort R, Trinidad Barnech G, Garat B, Duhagon M Noncoding RNA. 2023; 9(1).
PMID: 36827548 PMC: 9958706. DOI: 10.3390/ncrna9010015.
Carvajal-Moreno J, Hernandez V, Wang X, Li J, Yalowich J, Elton T J Pharmacol Exp Ther. 2022; 384(2):265-276.
PMID: 36410793 PMC: 9875313. DOI: 10.1124/jpet.122.001429.
Kang E, Jia Y, Wang J, Wang G, Chen H, Chen X CNS Neurosci Ther. 2022; 28(7):1081-1092.
PMID: 35481944 PMC: 9160452. DOI: 10.1111/cns.13845.
Huang H, Lin Y, Cui S, Huang Y, Tang Y, Xu J Nucleic Acids Res. 2021; 50(D1):D222-D230.
PMID: 34850920 PMC: 8728135. DOI: 10.1093/nar/gkab1079.