FRET-based dynamic structural biology: Challenges, perspectives and an appeal for open-science practices

Eitan Lerner, Anders Barth, Jelle Hendrix, Benjamin Ambrose, Victoria Birkedal, Scott C. Blanchard, Richard Börner, Hoi Sung Chung, Thorben Cordes, Timothy D. Craggs, Ashok A. Deniz, Jiajia Diao, Jingyi Fei, Ruben L. Gonzalez, Irina V. Gopich, Taekjip Ha, Christian A. Hanke, Gilad Haran, Nikos S. Hatzakis, Sungchul HohngSeok Cheol Hong, Thorsten Hugel, Antonino Ingargiola, Chirlmin Joo, Achillefs N. Kapanidis, Harold D. Kim, Ted Laurence, Nam Ki Lee, Tae Hee Lee, Edward A. Lemke, Emmanuel Margeat, Jens Michaelis, Xavier Michalet, Sua Myong, Daniel Nettels, Thomas Otavio Peulen, Evelyn Ploetz, Yair Razvag, Nicole C. Robb, Benjamin Schuler, Hamid Soleimaninejad, Chun Tang, Reza Vafabakhsh, Don C. Lamb, Claus A.M. Seidel, Shimon Weiss, Olga Boudker

Research output: Contribution to journalReview articlepeer-review

124 Citations (Scopus)


Single-molecule FRET (smFRET) has become a mainstream technique for studying biomolecular structural dynamics. The rapid and wide adoption of smFRET experiments by an ever- increasing number of groups has generated significant progress in sample preparation, measurement procedures, data analysis, algorithms and documentation. Several labs that employ smFRET approaches have joined forces to inform the smFRET community about streamlining how to perform experiments and analyze results for obtaining quantitative information on biomolecular structure and dynamics. The recent efforts include blind tests to assess the accuracy and the precision of smFRET experiments among different labs using various procedures. These multi-lab studies have led to the development of smFRET procedures and documentation, which are important when submitting entries into the archiving system for integrative structure models, PDB- Dev. This position paper describes the current ‘state of the art’ from different perspectives, points to unresolved methodological issues for quantitative structural studies, provides a set of ‘soft recommendations’ about which an emerging consensus exists, and lists openly available resources for newcomers and seasoned practitioners. To make further progress, we strongly encourage ‘open science’ practices.

Original languageEnglish
Article numbere60416
Publication statusPublished - 2021 Mar

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ASJC Scopus subject areas

  • General Neuroscience
  • General Biochemistry,Genetics and Molecular Biology
  • General Immunology and Microbiology


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