ArticleProtein science : a publication of the Protein Society2017
In vitro labeling strategies for in cellulo fluorescence microscopy of single ribonucleoprotein machines.
Article in Protein science : a publication of the Protein Society, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 20 citations in OpenAlex.
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- Surface-Tethering Enhances Precision in Measuring Diffusion Within 3D Protein Condensates.bioRxiv : the preprint server for biology · 2025Article
- BNP-Track: a framework for superresolved tracking.Nature methods · 2024Article
- Critical Assessment of Condensate Boundaries in Dual-Color Single Particle Tracking.The journal of physical chemistry. B · 2023Article
- Article
- DNA Tiling Enables Precise Acylation-Based Labeling and Control of mRNA.Angewandte Chemie (International ed. in English) · 2021Article
- Stealth Fluorescence Labeling for Live Microscopy Imaging of mRNA Delivery.Journal of the American Chemical Society · 2021Article
- Following the messenger: Recent innovations in live cell single molecule fluorescence imaging.Wiley interdisciplinary reviews. RNA · 2020Review
- Bioorthogonal mRNA labeling at the poly(A) tail for imaging localization and dynamics in live zebrafish embryos.Chemical science · 2020Article
- Establishing Preferred Product Characterization for the Evaluation of RNA Vaccine Antigens.Vaccines · 2019Review
- Coming Together: RNAs and Proteins Assemble under the Single-Molecule Fluorescence Microscope.Cold Spring Harbor perspectives in biology · 2019Review
- From "Cellular" RNA to "Smart" RNA: Multiple Roles of RNA in Genome Stability and Beyond.Chemical reviews · 2018Review
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
RNA plays a fundamental, ubiquitous role as either substrate or functional component of many large cellular complexes-"molecular machines"-used to maintain and control the readout of genetic information, a functional landscape that we are only beginning to understand. The cellular mechanisms for the spatiotemporal organization of the plethora of RNAs involved in gene expression are particularly poorly understood. Intracellular single-molecule fluorescence microscopy provides a powerful emerging tool for probing the pertinent mechanistic parameters that govern cellular RNA functions, including those of protein coding messenger RNAs (mRNAs). Progress has been hampered, however, by the scarcity of efficient high-yield methods to fluorescently label RNA molecules without the need to drastically increase their molecular weight through artificial appendages that may result in altered behavior. Herein, we employ T7 RNA polymerase to body label an RNA with a cyanine dye, as well as yeast poly(A) polymerase to strategically place multiple 2'-azido-modifications for subsequent fluorophore labeling either between the body and tail or randomly throughout the tail. Using a combination of biochemical and single-molecule fluorescence microscopy approaches, we demonstrate that both yeast poly(A) polymerase labeling strategies result in fully functional mRNA, whereas protein coding is severely diminished in the case of body labeling.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.