Journal ArticleDOI
Visualization of Single RNA Transcripts in Situ
Andrea M. Femino,Fredric S. Fay,Fredric S. Fay,Kevin E. Fogarty,Kevin E. Fogarty,Robert H. Singer,Robert H. Singer +6 more
TLDR
This approach extends the power of FISH to yield quantitative molecular information on a single cell by positioning probes along the transcription unit to determine the rates of transcription initiation and termination and messenger RNA processing.Abstract:
Fluorescence in situ hybridization (FISH) and digital imaging microscopy were modified to allow detection of single RNA molecules. Oligodeoxynucleotide probes were synthesized with five fluorochromes per molecule, and the light emitted by a single probe was calibrated. Points of light in exhaustively deconvolved images of hybridized cells gave fluorescent intensities and distances between probes consistent with single messenger RNA molecules. Analysis of beta-actin transcription sites after serum induction revealed synchronous and cyclical transcription from single genes. The rates of transcription initiation and termination and messenger RNA processing could be determined by positioning probes along the transcription unit. This approach extends the power of FISH to yield quantitative molecular information on a single cell.read more
Citations
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Nature, Nurture, or Chance: Stochastic Gene Expression and Its Consequences
TL;DR: Stochastic gene expression has important consequences for cellular function, being beneficial in some contexts and harmful in others, including the stress response, metabolism, development, the cell cycle, circadian rhythms, and aging.
Journal ArticleDOI
Gold nanoparticles for biology and medicine.
David A. Giljohann,Dwight S. Seferos,Dwight S. Seferos,Weston L. Daniel,Matthew D. Massich,Pinal C. Patel,Chad A. Mirkin +6 more
TL;DR: This Review highlights recent advances in the synthesis, bioconjugation, and cellular uses of gold nanoconjugates.
Journal ArticleDOI
Quantifying E. coli proteome and transcriptome with single-molecule sensitivity in single cells.
Yuichi Taniguchi,Paul J. Choi,Gene-Wei Li,Huiyi Chen,Mohan Babu,Jeremy Hearn,Andrew Emili,X. Sunney Xie +7 more
TL;DR: System-wide analyses of protein and mRNA expression in individual cells with single-molecule sensitivity using a newly constructed yellow fluorescent protein fusion library for Escherichia coli found that almost all protein number distributions can be described by the gamma distribution with two fitting parameters which, at low expression levels, have clear physical interpretations as the transcription rate and protein burst size.
Journal ArticleDOI
RNAscope: A Novel in Situ RNA Analysis Platform for Formalin-Fixed, Paraffin-Embedded Tissues
Fai Wang,John G. Flanagan,Nan Su,Li Chong Wang,Son Bui,Allissa Nielson,Xingyong Wu,Hong Thuy Vo,Xiao Jun Ma,Yuling Luo +9 more
TL;DR: RNAscope is described, a novel RNA ISH technology with a unique probe design strategy that allows simultaneous signal amplification and background suppression to achieve single-molecule visualization while preserving tissue morphology and may enable rapid development of RNAISH-based molecular diagnostic assays.
Journal ArticleDOI
Stochastic mRNA Synthesis in Mammalian Cells
TL;DR: The results demonstrate that gene expression in mammalian cells is subject to large, intrinsically random fluctuations and raise questions about how cells are able to function in the face of such noise.
References
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Journal ArticleDOI
Effect of protein synthesis inhibitors on growth factor activation of c-fos, c-myc, and actin gene transcription.
TL;DR: The results suggest that in PC12 cells c-fos transcription is activated by a protein-synthesis-independent mechanism, whereas c-myc stimulation requires new protein synthesis.
Journal ArticleDOI
Mating Type Switching in Yeast Controlled by Asymmetric Localization of ASH1 mRNA
Roy M. Long,Robert H. Singer,Robert H. Singer,Xiuhua Meng,Xiuhua Meng,Isabel Gonzalez,Isabel Gonzalez,Kim Nasmyth,Kim Nasmyth,Ralf-Peter Jansen,Ralf-Peter Jansen +10 more
TL;DR: It is shown that ASH1 messenger RNA (mRNA) preferentially accumulates in daughter cells by a process that is dependent on actin and myosin, and suggests that localization of mRNA may have been an early property of the eukaryotic lineage.
Journal ArticleDOI
Isoform-specific 3'-untranslated sequences sort alpha-cardiac and beta-cytoplasmic actin messenger RNAs to different cytoplasmic compartments.
TL;DR: It is proposed that the mechanism of mRNA localization facilitates actin isoform sorting in the cytoplasm and 3'UTR sequences play a key role in modulating the distribution of actin mRNAs in muscle cells.
Journal ArticleDOI
Transcription factor TFIID recruits factor CPSF for formation of 3′ end of mRNA
TL;DR: A link between transcription initiation and elongation by RNA polymerase II and processing of the 3′ end of mRNA is revealed and polyadenylation becomes less efficient due to incomplete assembly of TFIID on recombinant TBP.
Journal ArticleDOI
Nonrandom gene organization: structural arrangements of specific pre-mRNA transcription and splicing with SC-35 domains.
TL;DR: Results support a model reconciling light and electron microscopic observations which proposes that transcription of some specific genes occurs at the border of domains, which may also function in the assembly or distribution of RNA metabolic components.