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An Adaptable High-Throughput Technology Enabling the Identification of Specific Transcription Modulators

Artikel i vetenskaplig tidskrift
Författare T. Bergbrede
Emily Hoberg
N. G. Larsson
Maria Falkenberg
Claes M Gustafsson
Publicerad i Slas Discovery
Volym 22
Nummer/häfte 4
Sidor 378-386
ISSN 2472-5552
Publiceringsår 2017
Publicerad vid Institutionen för biomedicin
Sidor 378-386
Språk English
Länkar doi.org/10.1177/2472555217690326
Ämnesord Antiviral drugs, fluorescence methods, nucleic acid chemistry, assays, labeling, binding or purification, mitochondrial rna-polymerase, acute myeloid-leukemia, hepatitis-c virus, in-vitro, therapy, cancer, dna, pcr, Biochemistry & Molecular Biology, Biotechnology & Applied Microbiology, Chemistry
Ämneskategorier Biokemi och molekylärbiologi, Mikrobiologi, Kemi

Sammanfattning

Mitochondria harbor the oxidative phosphorylation (OXPHOS) system, which under aerobic conditions produces the bulk of cellular adenosine triphosphate (ATP). The mitochondrial genome encodes key components of the OXPHOS system, and it is transcribed by the mitochondrial RNA polymerase, POLRMT. The levels of mitochondrial transcription correlate with the respiratory activity of the cell. Therefore, compounds that can increase or decrease mitochondrial gene transcription may be useful for fine-tuning metabolism and could be used to treat metabolic diseases or certain forms of cancer. We here report the establishment of a novel high-throughput assay technology that has allowed us to screen a library of 430,000 diverse compounds for effects on mitochondrial transcription in vitro. Following secondary screens facilitated by the same assay principle, we identified 55 compounds that efficiently and selectively inhibit mitochondrial transcription and that are active also in cell culture. Our method is easily adaptable to other RNA or DNA polymerases and varying spectroscopic detection technologies.

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