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Characterizing single-molecule dynamics of viral RNA-dependent RNA polymerases with multiplexed magnetic tweezers

Journal Article (2022)
Author(s)

Louis Kuijpers (TU Delft - BN/Nynke Dekker Lab)

Theo van Laar (TU Delft - BN/Nynke Dekker Lab)

Richard Janissen (TU Delft - BN/Cees Dekker Lab)

Nynke H. Dekker (TU Delft - BN/Nynke Dekker Lab)

DOI related publication
https://doi.org/10.1016/j.xpro.2022.101606 Final published version
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Publication Year
2022
Language
English
Related content
Issue number
3
Volume number
3
Article number
101606
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271
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Abstract

Multiplexed single-molecule magnetic tweezers (MT) have recently been employed to probe the RNA synthesis dynamics of RNA-dependent RNA polymerases (RdRp). Here, we present a protocol for simultaneously probing the RNA synthesis dynamics of hundreds of single polymerases with MT. We describe the preparation of a dsRNA construct for probing single RdRp kinetics. We then detail the measurement of RdRp RNA synthesis kinetics using MT. The protocol is suitable for high-throughput probing of RdRp-targeting antiviral compounds for mechanistic function and efficacy. For complete details on the use and execution of this protocol, please refer to Janissen et al. (2021).