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The higher-order structural organization of the cell nucleus reflects the underlying genome-wide transcriptional activity and macromolecular transport processes. To study the microscopic organization of RNA distribution within the nucleus, a combinatorial library of fluorescent styryl molecules was synthesized and screened for an in vitro RNA response and live cell nuclear imaging. Four different cell lines (HeLa, A549, 3T3, and 3T3-L1) were analyzed in terms of higher-order nuclear organization. We identified RNA-selective dyes with better imaging properties relative to commercially available SYTORNASelect dye; the selected dyes were also cell permeant, photostable, and well tolerated by the cells. Our dyes also had very good counterstain compatibility with Hoechst and DAPI, which could help to image the DNA distribution in relation to RNA distribution in live cells and therefore reveal different patterns of RNA-DNA colocalization.
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http://dx.doi.org/10.1016/j.chembiol.2006.04.007 | DOI Listing |
PLoS Comput Biol
September 2025
Department of Evolutionary Biology and Environmental Studies, University of Zurich, Zurich, Switzerland.
Ubiquity of cancer across the tree of life yields opportunities to understand variation in cancer defences across species. Peto's paradox, the finding that large-bodied species do not suffer from more cancer despite having more cells at risk of oncogenic mutations compared to small species, can be explained if large size selects for better cancer defences. Since birds live longer than non-flying mammals of equivalent size, and are descendants of moderate-sized dinosaurs, we ask whether ancestral cancer defences are retained if body size shrinks in a lineage.
View Article and Find Full Text PDFSTAR Protoc
September 2025
Laboratory of Genome Integrity, CCR, NCI, NIH, Bethesda, MD, USA. Electronic address:
Tracking the translocation of fluorescent-based reporters at the single-cell level in living mouse embryos requires specialized expertise in mouse embryology and deep computational skills. Here, we detail an approach to quantify cyclin-dependent kinase (CDK) activity levels in single cells throughout different stages of the pre-implantation embryo. We discuss in vitro culture strategies that enable efficient live fluorescent confocal image acquisition and subsequent cell tracking.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
September 2025
Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur 208016, India.
Agonist-induced interaction of G protein-coupled receptors (GPCRs) with β-arrestins (βarrs) is a critical mechanism that regulates the spatiotemporal pattern of receptor localization and signaling. While the underlying mechanism governing GPCR-βarr interaction is primarily conserved and involves receptor activation and phosphorylation, there are several examples of receptor-specific fine-tuning of βarr-mediated functional outcomes. Considering the key contribution of conformational plasticity of βarrs in driving receptor-specific functional responses, it is important to develop novel sensors capable of reporting distinct βarr conformations in cellular context.
View Article and Find Full Text PDFProbiotics Antimicrob Proteins
September 2025
School of Life Science, Liaoning Normal University, Dalian, 116081, China.
Cutibacterium acnes (C. acnes, formerly classified as Propionibacterium acnes) is a Gram-positive bacterium that contributes to the development of acne vulgaris, resulting in inflammation and pustule formation on the skin. In this study, we developed and synthesized a series of antimicrobial peptides (AMPs) that are derived from the skin secretion of Rana chensinensis.
View Article and Find Full Text PDFJ Cell Biol
November 2025
Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Phosphatidic acid (PA) regulates lipid homeostasis and vesicular trafficking, yet high-affinity tools to study PA in live cells are lacking. We identified the lipin-like sequence of Nir1 (PILS-Nir1) as a candidate PA biosensor based on structural analysis of Nir1's LNS2 domain. Using liposome-binding assays and pharmacological and genetic manipulations in HEK293A cells expressing fluorescent PILS-Nir1, we found that while PILS-Nir1 binds PA and PIP2in vitro, only PA is necessary and sufficient for membrane localization in cells.
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