ERYTHROPOIESIS STUDY OF MIR-7132 IN ANTARTCTIC ICEFISH CHIONODRACO HAMATUS
microRNAs are small non-coding RNAs acting at post-transcriptional level by binding the target gene 3'UTR (untranslated region),and have an important effect on the erythropoiesis.Chionodraco hamatus is the only known vertebrates with a few non-functional erythrocytes.Previous findings showed that the highly upregulated expression of microRNAs in the head kidney of the animal could inhibit its erythropoiesis.By applying zebrafish microinjection,dual-luciferase reporter,and the target prediction method,we studied the erythropoiesis of miR-7132 that highly expressed in the head kidney.Results show that,after being injected with miR-7132,the expression level of hemoglobin in zebrafish embryos dramatically reduced as observed after o-dianisidine staining,indicating that the overexpression of miR-7132 repressed the erythropoiesis process in zebrafish embryos.The 3'UTR of ALAS2 (i.e.,5'-aminolevulinate synthase 2),the limited enzymes of hemoglobin production,was obtained from transcriptome sequencing data blasting unpublished icefish genome sequences.The dual-luciferase vector containing the 3'UTR of ALAS2 was constructed and co-transfected with miR-7132,and the activity of luciferase was detected.The expression vector containing the GFP (green fluorescent protein) fluorescent protein was constructed.Zebrafish embryos were then micro-injected with the mixture of miR-7132 and GFP plasmid,and the expression of GFP protein was detected by the western blotting.Therefore,miR-7132 could reduce significantly the relative activity of luciferase in 293T cells.The GFP fluorescent intensity could be decreased significantly when injected with miR-7132;and miR-7132 could regulate erythropoiesis in the Antarctic icefish by repressing the expression of ALAS2.
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万方数据
维普
