MicroRNAs (miRNAs) are short segments of non-coding RNAs that regulate the translation of messenger RNA (mRNA) into protein. Glycogenes, i.e., the genes that control glycosylation, are highly regulated by miRNAs. The glycan motif a-1,2 fucose is an important component of Lewisb and Lewisy and is the defining feature of the blood group H antigen. H antigen is the precursor of the ABO blood group antigens, if it is not present ABO blood group antigens cannot be formed. FUT1 and FUT2 genes encode a Golgi stack enzyme α-1,2-fucosyltransferase that catalyzes the transfer of 1,2-linked fucose to galactose residue of glycans. In this project, we aim to generate high throughput data that shows the regulation of FUT1 and FUT 2 by miRNA using miRFluR high-throughput assay. This assay utilized genetically encoded dual-color fluorescence reporters to identify regulatory miRNAs thus generating a comprehensive map of the regulation of FUT1 and FUT2 by approximately 2700 human miRNAs. pMIR-3’ UTR sensors of FUT1and FUT2 have been developed. From our high throughput assay, we have identified 76 regulatory miRNAs for FUT1. For these regulatory miRNAs, the data have been validated using western blot, lectin binding assay, and q-RT-PCR experiments in multiple cell lines. To summarize, this study contributes to the understanding of the miRNA regulations of FUT1 and FUT 2 and their significance for normal biological function.

Mapping the miRNA regulation of α-1,2 fucosylation

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ACS Fall 2022

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