zma-miR164a-5p: Zma-mir164a-5p is a type of microRNA that has been found to bind to the porcine target genes CSPG4, OTX1, and PLAGL2, suggesting that it can be absorbed from the diet and target specific genes [PMC7502231]. A dual-luciferase assay was performed to demonstrate the relationships between zma-mir164a-5p and these target genes [PMC5428504]. Through in silico analysis, 50 potential target porcine mRNAs for zma-mir164a-5p were predicted [PMC5428504]. Evidence from the miR-TRAP approach showed that zma-mir164a-5p can function similarly to endogenous miRNAs in porcine cells and target some of its predicted target genes in the Argonaute/RISC complex [PMC5428504]. The potential targeted mRNAs containing zma-mir164a-5p binding sites were synthesized for further analysis [PMC5428504]. qRT-PCR analysis revealed that 73.33% of the potential targets showed a significant increase in transcript levels, with 53.33% showing greater than 2-fold enrichment compared to control samples [PMC5428504]. The expression levels of zma-mir164a-5p and other maize-derived miRNAs were analyzed using qRT-PCR after feeding fresh maize to pigs, showing an increase in their abundance within 24 hours [PMC5428504]. Luciferase activity assays confirmed that zma-mir164a-5p significantly reduced luciferase activity for wild-type target genes but had no effect on mutant-type targets [PMC5428504]. TargetScan and NCBI Blast were used to predict potential target genes of zma-mir164a-5p in pigs, with their minimum free energy values evaluated by RNAhybrid [PMC5428504]. The Mir-TrAP System kit was used to identify specific microRNA targets in mammalian cells, including those targeted by zma-mir164a-5p [PMC5428504]. Additionally, zma-mir164a-5p was found to regulate six NAC genes [PMC9361504].
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