Induction of Translational Readthrough across the Thalassemia-Causing Premature Stop Codon in β-Globin-Encoding mRNA

Kar, D and Sellamuthu, K and Kumar, SD and Eswarappa, SM (2020) Induction of Translational Readthrough across the Thalassemia-Causing Premature Stop Codon in β-Globin-Encoding mRNA. In: Biochemistry, 59 (1). pp. 80-84.

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Abstract

Nonsense mutations that result in premature stop codons in the HBB gene cause β-thalassemia. This disease is characterized by a reduced hemoglobin level due to the lack of β-globin. Compounds that induce translational readthrough across the thalassemia-causing premature stop codon will have therapeutic benefits. Currently available molecules that induce translational readthrough lack specificity, and some of them show toxicity after prolonged use. In this study, we have developed an oligonucleotide-based approach to induce translational readthrough across the thalassemia-causing premature stop codon. Oligonucleotides that target HBB mRNA downstream of the premature stop codon could induce translational readthrough, generating a full-length β-globin protein. We show this effect using fluorescence- A nd luminescence-based readthrough assays and by Western blot. Remarkably, the amount of oligonucleotide-induced translational readthrough product is comparable to that of the protein generated by normal translation when there was no premature stop codon. Thus, these oligonucleotides, with certain modifications, have the potential to be used as drugs for the treatment of β-thalassemia. Also, this strategy can be extended to treat other genetic diseases caused by premature stop codons. © 2019 American Chemical Society.

Item Type:	Journal Article
Publication:	Biochemistry
Publisher:	American Chemical Society
Additional Information:	The copyright for this article belongs to American Chemical Society.
Keywords:	Oligonucleotides; Proteins; Signal encoding, Fluorescence and luminescence; Genetic disease; Hemoglobin levels; Stop codons; Therapeutic benefits; Western blots, Hemoglobin, antisense oligonucleotide; argonaute 1 protein; fusion protein; hemoglobin beta chain; messenger RNA; Renilla luciferin 2 monooxygenase; vasculotropin A; antisense oligonucleotide; hemoglobin beta chain; messenger RNA; oligodeoxyribonucleotide, Article; binding site; cystic fibrosis; DNA modification; Duchenne muscular dystrophy; flow cytometry; fluorescence; fluorescence microscopy; gene location; gene mutation; gene targeting; gene translocation; genetic transfection; glycation; HBB gene; hemophilia; human; luciferase assay; luminescence; molecular weight; priority journal; real time polymerase chain reaction; ribosome; spinal muscular atrophy; stop codon; thalassemia; Western blotting; drug effect; genetics; HEK293 cell line; metabolism; protein synthesis; stop codon; thalassemia, beta-Globins; Codon, Terminator; HEK293 Cells; Humans; Oligodeoxyribonucleotides; Oligonucleotides, Antisense; Protein Biosynthesis; RNA, Messenger; Thalassemia
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	06 Feb 2023 10:00
Last Modified:	06 Feb 2023 10:00
URI:	https://eprints.iisc.ac.in/id/eprint/79934

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