Filter
Associated Lab
- Aguilera Castrejon Lab (1) Apply Aguilera Castrejon Lab filter
- Ahrens Lab (1) Apply Ahrens Lab filter
- Betzig Lab (6) Apply Betzig Lab filter
- Feliciano Lab (1) Apply Feliciano Lab filter
- Gonen Lab (1) Apply Gonen Lab filter
- Hess Lab (2) Apply Hess Lab filter
- Keller Lab (1) Apply Keller Lab filter
- Lavis Lab (15) Apply Lavis Lab filter
- Lippincott-Schwartz Lab (12) Apply Lippincott-Schwartz Lab filter
- Remove Liu (Zhe) Lab filter Liu (Zhe) Lab
- O'Shea Lab (1) Apply O'Shea Lab filter
- Podgorski Lab (1) Apply Podgorski Lab filter
- Schreiter Lab (1) Apply Schreiter Lab filter
- Singer Lab (2) Apply Singer Lab filter
- Stringer Lab (1) Apply Stringer Lab filter
- Svoboda Lab (1) Apply Svoboda Lab filter
- Tillberg Lab (1) Apply Tillberg Lab filter
- Tjian Lab (7) Apply Tjian Lab filter
- Turner Lab (1) Apply Turner Lab filter
Associated Project Team
Associated Support Team
- Anatomy and Histology (1) Apply Anatomy and Histology filter
- Electron Microscopy (2) Apply Electron Microscopy filter
- Integrative Imaging (2) Apply Integrative Imaging filter
- Molecular Genomics (2) Apply Molecular Genomics filter
- Primary & iPS Cell Culture (3) Apply Primary & iPS Cell Culture filter
- Quantitative Genomics (1) Apply Quantitative Genomics filter
- Scientific Computing Software (1) Apply Scientific Computing Software filter
- Viral Tools (1) Apply Viral Tools filter
- Vivarium (1) Apply Vivarium filter
Publication Date
53 Janelia Publications
Showing 51-53 of 53 resultsOur ability to unambiguously image and track individual molecules in live cells is limited by packing of multiple copies of labeled molecules within the resolution limit. Here we devise a universal genetic strategy to precisely control copy number of fluorescently labeled molecules in a cell. This system has a dynamic titration range of >10,000 fold, enabling sparse labeling of proteins expressed at different abundance levels. Combined with photostable labels, this system extends the duration of automated single-molecule tracking by 2 orders of magnitude. We demonstrate long-term imaging of synaptic vesicle dynamics in cultured neurons as well as in intact zebrafish. We found axon initial segment utilizes a "waterfall" mechanism gating synaptic vesicle transport polarity by promoting anterograde transport processivity. Long-time observation also reveals that transcription factor hops between clustered binding sites in spatially-restricted sub-nuclear regions, suggesting that topological structures in the nucleus shape local gene activities by a sequestering mechanism. This strategy thus greatly expands the spatiotemporal length scales of live-cell single-molecule measurements, enabling new experiments to quantitatively understand complex control of molecular dynamics in vivo.
The assembly of sequence-specific enhancer-binding transcription factors (TFs) at cis-regulatory elements in the genome has long been regarded as the fundamental mechanism driving cell type-specific gene expression. However, despite extensive biochemical, genetic, and genomic studies in the past three decades, our understanding of molecular mechanisms underlying enhancer-mediated gene regulation remains incomplete. Recent advances in imaging technologies now enable direct visualization of TF-driven regulatory events and transcriptional activities at the single-cell, single-molecule level. The ability to observe the remarkably dynamic behavior of individual TFs in live cells at high spatiotemporal resolution has begun to provide novel mechanistic insights and promises new advances in deciphering causal-functional relationships of TF targeting, genome organization, and gene activation. In this review, we review current transcription imaging techniques and summarize converging results from various lines of research that may instigate a revision of models to describe key features of eukaryotic gene regulation.
YAP/TEAD signaling is essential for organismal development, cell proliferation, and cancer progression. As a transcriptional coactivator, how YAP activates its downstream target genes is incompletely understood. YAP forms biomolecular condensates in response to hyperosmotic stress, concentrating transcription-related factors to activate downstream target genes. However, whether YAP forms condensates under other signals, how YAP condensates organize and function, and how YAP condensates activate transcription in general are unknown. Here, we report that endogenous YAP forms sub-micron scale condensates in response to Hippo pathway regulation and actin cytoskeletal tension. YAP condensates are stabilized by the transcription factor TEAD1, and recruit BRD4, a coactivator that is enriched at active enhancers. Using single-particle tracking, we found that YAP condensates slowed YAP diffusion within condensate boundaries, a possible mechanism for promoting YAP target search. These results reveal that YAP condensate formation is a highly regulated process that is critical for YAP/TEAD target gene expression.