4.8 Article

Actin chromobody imaging reveals sub-organellar actin dynamics

Journal

NATURE METHODS
Volume 17, Issue 9, Pages 917-+

Publisher

NATURE PORTFOLIO
DOI: 10.1038/s41592-020-0926-5

Keywords

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Funding

  1. Waitt Foundation
  2. Core Grant application National Cancer Institute (NCI) Cancer Center Support Grant (CCSG) [CA014195]
  3. Transgenic Core Facility of the Salk [014195]
  4. Human Frontier Science Program (HFSP) [RGP0021/2016]
  5. Cluster of Excellence Centre for Integrative Biological Signaling Studies (CIBSS) [EXC-2189]
  6. NIH [R01 AR069876]
  7. Salk Institute Audrey Geisel Chair in Biomedical Science
  8. National Institute of General Medical Sciences (NIGMS) [R15 GM119077]
  9. University of Richmond School of Arts Sciences

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The actin cytoskeleton plays multiple critical roles in cells, from cell migration to organelle dynamics. The small and transient actin structures regulating organelle dynamics are challenging to detect with fluorescence microscopy, making it difficult to determine whether actin filaments are directly associated with specific membranes. To address these limitations, we developed fluorescent-protein-tagged actin nanobodies, termed 'actin chromobodies' (ACs), targeted to organelle membranes to enable high-resolution imaging of sub-organellar actin dynamics. Genetically encoded fluorescent-protein-tagged actin nanobodies fused to organelle membrane-targeting sequences enables high-resolution imaging of transient organelle-actin contact sites in live cells.

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