CRISPR/Cas9 Ablated BCL11A Unveils the Genes with Possible Role of Globin Switching

Abstract

Purpose: HbF upregulation is a mitigating factor in β-hemoglobinopathies therapy like β-thalassemia and sickle cell diseases. Finding molecular mechanisms and the key regulators responsible for globin switching could be helpful to develop effective ways to HbF upregulation. In our paper published recently, we introduced certain factors probably responsible for globin switching using in-silico study. The aim of this paper is experimental confirmation of the factors. Methods: We stablished K562 cell line with BCL11A knock down leading to increase in HBG1/2 using CRISPR/Cas9 system. After that, we determined the expression of factors introduced in previous paper by qPCR. Results: Results showed that BCL11A was substantially downregulated and HBG1/2 was upregulated in transfected K562 cells. Also, the experimental and bioinformatics data were consistent with each other for three genes, HIST1H2BL، TRIM58، AL13243.2. These data Conclusion: BCL11A is a good candidate for the treatment of β-hemoglobinopathies, with high HbF reactivation. In addition, HIST1H2BL, TRIM58 and AL13243.2 probably play a key role in the mechanism of hemoglobin switching. To further validate the selected genes, more experimental in-vivo and in-vitro studies are required.