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

Louis Kuijpers, Theo van Laar, Richard Janissen*, Nynke H. Dekker

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

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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).

Original languageEnglish
Article number101606
JournalSTAR Protocols
Volume3
Issue number3
DOIs
Publication statusPublished - 2022

Keywords

  • Biophysics
  • Microbiology
  • Molecular biology
  • Molecular/Chemical probes
  • Single-molecule assays

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