Functional mechanism of miR-30e on hematopoiesis in zebrafish
PEI Tianying1, YAN Shuaishuai1, XU Qianghua1,2,3,4*
1.College of Marine Sciences, Shanghai Ocean University, Shanghai 201306, China； 2.Key Laboratory of Sustainable Exploitation of Ocean Fisheries Resources, Ministry of Education, Shanghai 201306, China； 3.National Distant-water Fisheries Engineering Research Center, Shanghai 201306, China； 4.Collaborative Innovation Center for Distant-water Fisheries, Shanghai 201306, China
Abstract: MicroRNA (miRNA) as a type of single-stranded RNA molecule encoded by endogenous genes is involved in the regulation of post-transcriptional gene expression and plays an important role in hematopoiesis. In order to study the mechanism of miR-30e on hematopoiesis in zebrafish Daniorerio, the target gene of zebrafish miR-30e was predicted by bioinformatic software, based on the significantly highly expressed miR-30e in the blood tissue of the Qinghai-Tibetan plateau schizothoracine fish, and a double fluorescence plasmid containing zebrafish ALAS2-3′UTR sequence was constructed. The reporter plasmid was co-transfected with zebrafish miR-30e, and cell luciferase activity was subsequently examined. Meanwhile, green fluorescent protein particles containing zebrafish ALAS2-3′UTR sequence were established, co-micro-injected with miR-30e into zebrafish embryos, and the GFP fluorescence intensity and protein expression changes were subsequently observed. The results showed that the expression of hemoglobin was significantly reduced in the zebrafish embryos micro-injected with miR-30e mimics 48 hours after micro-injection, the relative activity of luciferase in the 293T cells transfected with miR-30e was significantly reduced (P<0.05). Moreover, the GFP fluorescence intensity and the GFP protein expression in the zebrafish embryos micro-injected with miR-30e mimics 24 hours after micro-injection were significantly reduced. It is concluded that ALAS2 is one of the target genes of miR-30e which inhibits erythropoiesis by reducing the expression levels of ALAS2.
2021, Vol. 36 
