miRNA expression and interaction with the 3'UTR of FMR1 in FRAXopathy pathogenesis

Alexander A Dolskiy; Andrey A Yarushkin; Irina V Grishchenko; Natalya A Lemskaya; Alexey V Pindyurin; Lidiya V Boldyreva; Vladimir O Pustylnyak; Dmitry V Yudkin

doi:10.1016/j.ncrna.2020.11.006

miRNA expression and interaction with the 3'UTR of FMR1 in FRAXopathy pathogenesis

Noncoding RNA Res. 2020 Dec 3;6(1):1-7. doi: 10.1016/j.ncrna.2020.11.006. eCollection 2021 Mar.

Authors

Alexander A Dolskiy¹, Andrey A Yarushkin^{2

3}, Irina V Grishchenko¹, Natalya A Lemskaya^{1

4}, Alexey V Pindyurin⁴, Lidiya V Boldyreva⁴, Vladimir O Pustylnyak^{2

3}, Dmitry V Yudkin¹

Affiliations

¹ State Research Center of Virology and Biotechnology "Vector", Federal Service for Surveillance on Consumer Rights Protection and Human Well-being (FBRI SRC VB "Vector", Rospotrebnadzor), Koltsovo, Novosibirsk Region, Russia.
² Federal Research Center of Fundamental and Translational Medicine, Novosibirsk, Novosibirsk Region, Russia.
³ Novosibirsk State University, Novosibirsk, Novosibirsk Region, Russia.
⁴ Institute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Novosibirsk Region, Russia.

Abstract

FRAXopathies are caused by the expansion of the CGG repeat in the 5'UTR of the FMR1 gene, which encodes the protein responsible for the synthesis of FMRP. This mutation leads to dramatic changes in FMRP expression at both the mRNA and protein levels. Evidence is emerging that changes in FMR1 mRNA expression can lead to the dysregulation of the miRNAs that target its 3'UTR. In the present work, B-lymphocyte cell lines obtained from patients with FRAXopathies were used, and a wide variety of FMR1 gene activities were observed, allowing the identification of the relationships between FMR1 dysregulation and miRNA activity. We studied the expression levels of eight miRNAs that target the FMR1 gene. To prove the interaction of the studied miRNAs with FMR1, a plasmid was constructed that possesses three primary structures: the miRNA gene, with expression driven by an inducible promoter; a constitutively expressed FusionRed reporter; and an eGFP reporter followed by the 3'UTR of the FMR1 gene. We evaluated changes in miRNA expression in response to alterations in FMR1 gene activity in a model cell line as well as interactions with some miRNAs with the FMR1 3'UTR.

Keywords: 3′UTR; FMR1; Fragile X associated tremor/ataxia syndrome; Fragile X syndrome; Fragile X-associated primary ovarian insufficiency; miRNAs.