Expression analysis of gamma globin gene switching associated micro RNAs, flowing BCL11A gene inhibition

Background and Objective :MicroRNAs are small, about 18 to 25 nucleotides, single-stranded RNA molecules that are targeting between 10% and 30% of the human genome. MicroRNA can target several genes, and also a gene that binds to several miRNAs. By connecting them to targets gene, the silencing occur after the translation.The siRNA (small or short / silecing interfering RNA) or RNAi is a synthetic RNA at two-strand structure. The appropriate length is between 19-21 nucleotides. The siRNA specifically targets the mRNA and inducing silencing in protein coding genes similar to the microRNA, the degradation of the target mRNA results in non-translation. MiR- 15a, miR- 16, miR-486-3p, miR- 23a and miR-27a play an important role in the hematopoiesis, especially in switching of the gamma to beta gene. Materials and Methods: The aim of this study is to investigate changes in the expression of miRNAs before and after inhibiting the gamma-beta gene switch by siRNA against the BCL11A gene. For this reason, peripheral blood mononuclear cells that contain stem cells are isolated by Ficoll-Paque.They were culture in the IMDM medium containing supplementary materials for differentiation to erythroid series for a period of 28 days and stored in a 37-degree incubator containing 5% CO2. Every 2-3 days, identification of the cells was done by two methods: Staining with Geimsa of large erythroid colonies and flow cytometry for CD235a marker.On the 21st day, cells were introduced by insertion of siRNA against BCL11A. Result: The results were evaluated using REST software. The data show that changes in the miRNAs examined before and after the switching of the gamma gene to the beta are significant. The results of this study can suggest new therapeutic approaches for patients with hemoglobinopathy.

کلیدواژه ها:

BCL11A ، miRNAs ، siRNA ، Hemoglobinopathies