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Updated: Aug 6, 2025

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
Published on: June 12, 2018
MicroRNA sequence variation can impact interactions with target mRNA in cattle
Dan Hao1, Xiao Wang2, Yu Yang3
1Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus C, Denmark; College of Animal Science and Technology, Northwest A&F University, Shaanxi Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, 712100 Yangling, Shaanxi, China.
Background:
Small non-coding microRNAs (miRNAs) are important modulators at post-transcriptional levels. Single-nucleotide polymorphisms (SNPs) located in miRNA genes can alter the secondary structure of pre-miRNA to either impair or promote the miRNA maturation processes. Furthermore, SNPs located in the miRNA seed regions can stabilize or disturb miRNA-target interactions, thereby, quantitatively influence the expression of target genes. Therefore, the main objective of this study was to detect SNPs in bovine miRNAs using the whole-genome re-sequencing datasets of 1632 cattle of five breeds from the 1000 bull genomes project.
Results:
In total, our study identified 1109, 334, and 130 SNPs in the miRNA precursor, mature, and seed regions, respectively. The heterozygosity values were generally less than 0.3, and the minor allele frequencies (MAFs) were mainly less than 0.1. Most SNPs were in Hardy-Weinberg equilibrium (HWE) (HWE-P > 0.05). Furthermore, we found that the majority of SNPs (MAF > 0.1 and HWE-P > 0.05) in the miRNA seed regions altered the repertoire of target genes, which in turn were enriched in different KEGG pathways or GO terms. Thus target prediction for bta-miR-2888 revealed loss of 309 target genes and gain of 691 target genes. The 691 gained target genes were significantly enriched in 60 KEGG pathways and 21 GO terms.
Conclusion:
In summary, our study identified candidate SNPs in miRNA precursor, mature, and seed regions that are likely to affect target RNA interactions, thereby potentially influencing cattle phenotypic traits.
Insights
Single-nucleotide polymorphisms (SNPs) in bovine microRNAs (miRNAs) were identified, potentially impacting gene regulation and cattle traits. These genetic variations in miRNA seed regions alter target gene interactions and associated biological pathways.
Area of Science:
- Genomics
- Molecular Biology
- Bioinformatics
Background:
- MicroRNAs (miRNAs) are small non-coding RNAs regulating gene expression post-transcriptionally.
- Single-nucleotide polymorphisms (SNPs) in miRNA genes can affect miRNA maturation and target interactions.
- SNPs in miRNA seed regions can quantitatively influence target gene expression.
Purpose of the Study:
- To detect single-nucleotide polymorphisms (SNPs) in bovine microRNAs (miRNAs).
- To analyze the impact of these SNPs on miRNA maturation and target gene interactions.
- To utilize whole-genome re-sequencing data from the 1000 Bull Genomes Project.
Main Methods:
- Whole-genome re-sequencing data analysis of 1632 cattle from five breeds.
- Identification and characterization of SNPs in miRNA precursor, mature, and seed regions.
- Target gene prediction and enrichment analysis (KEGG pathways, GO terms).
Main Results:
- 1109, 334, and 130 SNPs identified in miRNA precursor, mature, and seed regions, respectively.
- Most identified SNPs were in Hardy-Weinberg equilibrium with low heterozygosity and minor allele frequencies.
- SNPs in seed regions significantly altered target gene repertoires, with bta-miR-2888 showing gain/loss of numerous targets enriched in specific pathways.
Conclusions:
- Candidate SNPs in bovine miRNA regions were identified.
- These SNPs are likely to affect miRNA-target interactions.
- The identified SNPs may influence cattle phenotypic traits.
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