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| miR-122 Targets to Regulate BZW2 Gene Expression in Chicken(Gallus gallus) |
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Abstract MicroRNAs are a class of tiny, single-stranded noncoding RNAs that can target mRNA via base pairing and lead to translational repression or mRNA degradation at the level of gene posttranscription, which are involved in organ development, cell differentiation, proliferation, apoptosis and other biological processes, closely related with many diseases. Basic leucine zipper and W2 domains 2 (BZW2) functions in protein synthesis metabolism. The purpose of this study was to validate whether miR-122 could inhibit BZW2 gene expression in chicken (Gallus gallus). The bioinformatics method was used to investigate whether miR-122 could pair chicken BZW2 3'-UTR, furthermore dual-luciferase report system and mutant assay were used to validate that miR-122 repressed BZW2 expression. qRT-PCR was used to detect the expression level of BZW2 in leghorn hepatocellar (LHM) cells transfected with miRNA-122mimic and primary chicken hepatocytes transfected with LNA-antimiR-122. Bioinformatics prediction and dual luciferase report system proved that BZW2 was target gene of miR-122, target site mutation assay showed the complementary site of 3'UTR with miR-122 seeds region were target site of miR-122. After overexpression of miR-122 in leghorn hepatocellar (LMH) cells, BZW2 mRNA expression level decreased obviously. Using LNA-antimiR-122 to knockdown the miR-122 expression in primary chicken hepatocytes, the target gene BZW2 mRNA expression level increased significantly. These data suggested that BZW2 gene was target gene of miR-122 and BZW2 mRNA was negatively regulated by miR-122 in chicken hepatocytes. It will provide scientific basis on revealing a wide range of functions in the liver of miR-122 in chicken.
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Received: 12 July 2016
Published: 01 October 2016
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| Fund:National Natural Science Foundation of China |
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