Abstract
RNA turnover is an essential part of the gene expression pathway, and there are several experimental approaches for its determination. High-throughput measurement of global RNA turnover rates can provide valuable information about conditions or proteins that impact gene expression. Here, we present a protocol for mitochondrial RNA turnover analysis which involves metabolic labeling of RNA coupled with quantitative high-throughput fluorescent microscopy. This approach gives an excellent opportunity to discover new factors involved in mitochondrial gene regulation when combined with loss-of-function screening strategy.
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Acknowledgments
Studies were supported by the National Science Centre, Poland [UMO-2014/12/W/NZ1/00463 to R.J.S.]. Experiments were carried out with the use of CePT infrastructure financed by the European Union: the European Regional Development Fund (Innovative economy 2007–13, Agreement POIG.02.02.00-14-024/08-00).
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Kotrys, A.V., Borowski, L.S., Szczesny, R.J. (2021). High-Throughput Measurement of Mitochondrial RNA Turnover in Human Cultured Cells. In: Minczuk, M., Rorbach, J. (eds) Mitochondrial Gene Expression. Methods in Molecular Biology, vol 2192. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0834-0_11
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DOI: https://doi.org/10.1007/978-1-0716-0834-0_11
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