Quantitative RNA imaging in single live cells reveals age-dependent asymmetric inheritance

• Published in: Cell reports (Cambridge) Vol. 41; no. 7; p. 111656
• Main Authors: Kukhtevich, Igor V., Rivero-Romano, Mariana, Rakesh, Namisha, Bheda, Poonam, Chadha, Yagya, Rosales-Becerra, Paulina, Hamperl, Stephan, Bureik, Daniela, Dornauer, Scarlett, Dargemont, Catherine, Kirmizis, Antonis, Schmoller, Kurt M., Schneider, Robert
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.11.2022
• Subjects: aging; Cell Division; diSpinach aptamer; Inheritance Patterns; microfluidics; RNA; RNA aptamer; RNA imaging; RNA inheritance; S. cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins - genetics; single cells; yeast; RNA aptamer; RNA imaging; diSpinach aptamer; CP: Cell biology; RNA inheritance; single cells; aging; microfluidics; yeast; S. cerevisiae
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2022.111656

Asymmetric inheritance of cellular content through cell division plays an important role in cell viability and fitness. The dynamics of RNA segregation are so far largely unaddressed. This is partly due to a lack of approaches to follow RNAs over multiple cellular divisions. Here, we establish an approach to quantify RNA dynamics in single cells across several generations in a microfluidics device by tagging RNAs with the diSpinach aptamer. Using S. cerevisiae as a model, we quantitatively characterize intracellular RNA transport from mothers into their buds. Our results suggest that, at cytokinesis, ENO2 diSpinach RNA is preferentially distributed to daughters. This asymmetric RNA segregation depends on the lifespan regulator Sir2 and decreases with increasing replicative age of mothers but does not result from increasing cell size during aging. Overall, our approach opens more opportunities to study RNA dynamics and inheritance in live budding yeast at the single-cell level. [Display omitted] •diSpinach aptamer tagging to study RNA inheritance in cells on a microfluidics device•ENO2 RNA, visualized through diSpinach, is asymmetrically distributed to daughters•Loss of asymmetric RNA inheritance is a direct consequence of cellular aging•Sir2 is essential for the asymmetric RNA inheritance Kukhtevich et al. propose an approach to quantify RNA dynamics in single cells across several generations in a microfluidics device by tagging RNAs with the diSpinach aptamer. The results suggest that, in S. cerevisiae, ENO2 diSpinach RNA is inherited asymmetrically by daughters and that this is an aging-dependent phenomenon.