ArticleMolecular biology of the cell2014
Measuring fast gene dynamics in single cells with time-lapse luminescence microscopy.
Article in Molecular biology of the cell, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 24 citations in OpenAlex.
- A system of reporters for comparative investigation of EJC-independent and EJC-enhanced nonsense-mediated mRNA decay.Nucleic acids research · 2024Article
- Multidimensional characterization of inducible promoters and a highly light-sensitive LOV-transcription factor.Nature communications · 2023Article
- Tuning of Gene Expression inACS synthetic biology · 2022Article
- A transcriptional biosensor to monitor single cancer cell therapeutic responses by bioluminescence microscopy.Theranostics · 2022Article
- Reporter gene comparison demonstrates interference of complex body fluids with secreted luciferase activity.Scientific reports · 2021Article
- In Vivo Simultaneous Analysis of Gene Expression by Dual-Color Luciferases inInternational journal of molecular sciences · 2020Article
- Capturing and Understanding the Dynamics and Heterogeneity of Gene Expression in the Living Cell.International journal of molecular sciences · 2020Review
- Glioblastoma Cell Resistance to EGFR and MET Inhibition Can Be Overcome via Blockade of FGFR-SPRY2 Bypass Signaling.Cell reports · 2020Article
- Using time-lapse fluorescence microscopy to study gene regulation.Methods (San Diego, Calif.) · 2019Review
- A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo.Nature biotechnology · 2016Article
- Real-time quantification of protein expression at the single-cell level via dynamic protein synthesis translocation reporters.Nature communications · 2016Article
- Different Mechanisms Confer Gradual Control and Memory at Nutrient- and Stress-Regulated Genes in Yeast.Molecular and cellular biology · 2015Article
- Tracking transience: a method for dynamic monitoring of biological events in Arabidopsis thaliana biosensors.Planta · 2015Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Time-lapse fluorescence microscopy is an important tool for measuring in vivo gene dynamics in single cells. However, fluorescent proteins are limited by slow chromophore maturation times and the cellular autofluorescence or phototoxicity that arises from light excitation. An alternative is luciferase, an enzyme that emits photons and is active upon folding. The photon flux per luciferase is significantly lower than that for fluorescent proteins. Thus time-lapse luminescence microscopy has been successfully used to track gene dynamics only in larger organisms and for slower processes, for which more total photons can be collected in one exposure. Here we tested green, yellow, and red beetle luciferases and optimized substrate conditions for in vivo luminescence. By combining time-lapse luminescence microscopy with a microfluidic device, we tracked the dynamics of cell cycle genes in single yeast with subminute exposure times over many generations. Our method was faster and in cells with much smaller volumes than previous work. Fluorescence of an optimized reporter (Venus) lagged luminescence by 15-20 min, which is consistent with its known rate of chromophore maturation in yeast. Our work demonstrates that luciferases are better than fluorescent proteins at faithfully tracking the underlying gene expression.
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