A Foundational Study for Normal F8 -Containing Mouse Models for the miRNA Regulation of Hemophilia A: Identification and Analysis of Mouse miRNAs that Downregulate the Murine F8 Gene

• Published in: International journal of molecular sciences 2020-08, Vol.21 (16), p.5621
• Main Authors: Jankowska, Katarzyna I, Chattopadhyay, Maitreyi, Sauna, Zuben E, Atreya, Chintamani D
• Format: Article
• Language: English
• Subjects: 3' Untranslated Regions - genetics; Animal models; Animals; Base Sequence; BASIC BIOLOGICAL SCIENCES; Binding sites; bleeding disorders; Coagulation; Coagulation factor VIII; Coagulation factors; Disease Models, Animal; Down-Regulation - genetics; Ectopic expression; Factor VIII - genetics; Factor VIII deficiency; Gene expression; Gene regulation; Genotypes; HEK293 Cells; HeLa Cells; Hemophilia; hemophilia A; Hemophilia A - genetics; Humans; Mammalian cells; Mice; MicroRNAs; microRNAs (miRNAs); MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; mouse model; mRNA; Mutation; Plasmids; Proteins; Reagents; Reporter gene; Software
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21165621

Hemophilia A (HA) is associated with defects in the gene, encoding coagulation factor VIII (FVIII). Our previous studies show that F8-targeting micro RNAs (miRNAs), a group of small RNAs involved in gene regulation, can downregulate F8 expression causing HA in individuals with normal F8-genotypes and increased HA severity in patients with mutations in . Understanding the mechanistic underpinnings of human genetic diseases caused or modulated by miRNAs require a small animal model, such as a mouse model. Here, we report a foundational study to develop such a model system. We identified the mouse 3'untranslated region (3'UTR) on murine -mRNA (mu -mRNA) that can bind to murine miRNAs. We then selected three miRNAs for evaluation: miR-208a, miR-351 and miR-125a. We first demonstrate that these three miRNAs directly target the 3'UTR of mu -mRNA and reduce the expression of a reporter gene (luciferase) mRNA fused to the mu -3' UTR in mammalian cells. Furthermore, in mouse cells that endogenously express the gene and produce FVIII protein, the ectopic expression of these miRNAs downregulated -mRNA and FVIII protein. These results provide proof-of-concept and reagents as a foundation for using a normal -containing mouse as a model for the miRNA regulation of normal in causing or aggravating the genetic disease HA.