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

• Published in: Gene reports Vol. 21; p. 100934
• Main Authors: Obeidi, Narges, Tamaddon, Gholamhossein, Ranjbaran, Reza, Khamisipour, Gholamreza, Saberi, Fatemeh
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01.12.2020
• Subjects: BCL11A; Gamma globin; Hemoglobinopathy; miRNAs; siRNA; dsRNA; Gamma globin; Hemoglobinopathy; FDA; IL; miRNAs; BCL11A; Epo; siRNA; KLF3; IMDM; SCA; FITC; Sp1; CPDA-1; FBS; miR; MYB; SCF; PCR
• ISSN: 2452-0144; 2452-0144
• DOI: 10.1016/j.genrep.2020.100934

MicroRNAs are small, single-stranded RNA molecules that are targeting between 10% and 30% of the human genome, also a gene that binds to several miRNAs. By connecting them to targets gene, the silencing occurs after the translation. The aim of this study was to investigate changes in the expression of miRNAs before and after inhibiting the gamma-beta gene switch by siRNA against the BCL11A gene. MiR- 15a, miR- 16, miR-486-3p, miR- 23a and miR-27a, important miRs in switching gamma to beta gene, were investigated. Peripheral blood mononuclear cells were isolated by Ficoll-Paque. They were cultured 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. On the 21st day, cells were introduced by insertion of siRNA against BCL11A. The expression of miR- 15a in the cultured cells gradually increased from the first day to the eighteenth day and decreased on 21 day. Expression pattern of miR- 16 was increased on the 8th day and decreased on the 22th day. MiR-23a decreased on day 8, increased on day 21, and remained unchanged on days 18 and 22. miR-27a increased the expression on day 8, decreased on day 21, and remained unchanged on days 18 and 22. Expression of miR-486-3p increased on the 8th, 18th and 21th days; also, on the 22thday, there was a decrease in expression of miR-486-3p. Our data showed that changes in the expression of miRNAs were significant. The results of this study suggest that inhibition of the gamma gene switch to beta, keeps hemoglobin F high, therefore, this could lead to improved symptoms in patients with hemoglobinopathy. •We aimed to assess changes in miRNAs expression before and after inhibiting γ-β gene switch by siRNA against the BCL11A gene.•The expression pattern of miR-15a, miR-16, miR-23a, miR-27a, and miR-486-3p were assessed on days 1, 8, 18, 21, and 22.•We found that inhibition of γ gene switch to β keeps Hb-F high.•This could lead to improved symptoms in patients with hemoglobinopathy.