20 min listen
hGRAD - a versatile 'one-fits-all' system for the acute depletion of RNA binding proteins in nuclear condensates
hGRAD - a versatile 'one-fits-all' system for the acute depletion of RNA binding proteins in nuclear condensates
ratings:
Length:
20 minutes
Released:
Aug 4, 2023
Format:
Podcast episode
Description
Link to bioRxiv paper:
http://biorxiv.org/cgi/content/short/2023.08.04.551933v1?rss=1
Authors: Muller-McNicoll, M., Zarnack, K., McNicoll, F., Keller, M., Sliskovic, I., Okuda, E. K., Riegger, R. J., Arnold, B.
Abstract:
Nuclear RNA binding proteins (RBPs) are difficult to study because they often belong to large protein families and form extensive networks of auto- and cross-regulation. They are highly abundant and often localize to condensates with a slow turnover, requiring long depletion times or knockouts that cannot distinguish between direct and indirect or compensatory effects. Here, we developed a system that is optimized for the rapid degradation of nuclear RBPs, called hGRAD. It comes as a 'one-fits-all' plasmid, and integration into any cell line that expresses endogenously GFP-tagged proteins allows an inducible, rapid and complete knockdown. We show that the nuclear RBPs SRSF3, SRSF5, SRRM2 and NONO are completely cleared from nuclear speckles and paraspeckles within two hours. hGRAD works in various cell types, is more efficient than other methods and does not require the expression of exogenous ubiquitin ligases. Combining SRSF5 hGRAD degradation with Nascent-seq uncovered highly dynamic transient transcript changes, compensatory mechanisms and that SRSF5 promotes transcript stability.
Copy rights belong to original authors. Visit the link for more info
Podcast created by Paper Player, LLC
http://biorxiv.org/cgi/content/short/2023.08.04.551933v1?rss=1
Authors: Muller-McNicoll, M., Zarnack, K., McNicoll, F., Keller, M., Sliskovic, I., Okuda, E. K., Riegger, R. J., Arnold, B.
Abstract:
Nuclear RNA binding proteins (RBPs) are difficult to study because they often belong to large protein families and form extensive networks of auto- and cross-regulation. They are highly abundant and often localize to condensates with a slow turnover, requiring long depletion times or knockouts that cannot distinguish between direct and indirect or compensatory effects. Here, we developed a system that is optimized for the rapid degradation of nuclear RBPs, called hGRAD. It comes as a 'one-fits-all' plasmid, and integration into any cell line that expresses endogenously GFP-tagged proteins allows an inducible, rapid and complete knockdown. We show that the nuclear RBPs SRSF3, SRSF5, SRRM2 and NONO are completely cleared from nuclear speckles and paraspeckles within two hours. hGRAD works in various cell types, is more efficient than other methods and does not require the expression of exogenous ubiquitin ligases. Combining SRSF5 hGRAD degradation with Nascent-seq uncovered highly dynamic transient transcript changes, compensatory mechanisms and that SRSF5 promotes transcript stability.
Copy rights belong to original authors. Visit the link for more info
Podcast created by Paper Player, LLC
Released:
Aug 4, 2023
Format:
Podcast episode
Titles in the series (100)
Metabolite profiling and cytotoxic activity of Andean potatoes: polyamines and glycoalkaloids as potential anticancer agents in human neuroblastoma cells in vitro by PaperPlayer biorxiv cell biology