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Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
Published on: August 6, 2014
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Quantitative RNA imaging in single live cells reveals age-dependent asymmetric inheritance
Igor V Kukhtevich1, Mariana Rivero-Romano2, Namisha Rakesh3
1Institute of Functional Epigenetics, Helmholtz Zentrum München, 85764 Neuherberg, Germany.
Cell Reports
|November 17, 2022
Summary
Researchers developed a new method to track RNA movement across cell divisions in budding yeast. They found that RNA is unequally distributed to daughter cells, influenced by aging and the Sir2 protein.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Asymmetric inheritance of cellular components is crucial for cell viability and fitness.
- RNA segregation dynamics during cell division remain largely uncharacterized due to limited tracking methods.
- Understanding RNA inheritance is key to comprehending cellular aging and function.
Purpose of the Study:
- To develop and apply a novel method for quantifying RNA dynamics across multiple cell generations.
- To investigate the mechanisms and factors influencing asymmetric RNA segregation in budding yeast.
- To characterize the relationship between RNA inheritance, cellular aging, and lifespan regulation.
Main Methods:
- Utilized a microfluidics device for long-term single-cell observation of Saccharomyces cerevisiae (budding yeast).
- Developed an RNA tagging system using the diSpinach aptamer for quantitative tracking of RNA molecules.
- Quantified intracellular RNA transport from mother to daughter cells across several generations.
Main Results:
- Demonstrated preferential distribution of ENO2 diSpinach RNA to daughter cells during cytokinesis.
- Showed that asymmetric RNA segregation is dependent on the lifespan regulator Sir2.
- Observed a decrease in RNA asymmetry with increasing replicative age of mother cells, independent of cell size.
Conclusions:
- The developed approach enables the study of RNA dynamics and inheritance in live budding yeast at the single-cell level.
- Asymmetric RNA segregation is a regulated process influenced by Sir2 and cellular age.
- Findings provide new insights into the role of RNA inheritance in cellular aging and fitness.
