Back to Search
Start Over
Quantitative photoconversion analysis of internal molecular dynamics in stress granules and other membraneless organelles in live cells.
- Source :
-
STAR protocols [STAR Protoc] 2020 Dec 10; Vol. 1 (3), pp. 100217. Date of Electronic Publication: 2020 Dec 10 (Print Publication: 2020). - Publication Year :
- 2020
-
Abstract
- Photoconversion enables real-time labeling of protein sub-populations inside living cells, which can then be tracked with submicrometer resolution. Here, we detail the protocol of comparing protein dynamics inside membraneless organelles in live HEK293T cells using a CRISPR-Cas9 PABPC1-Dendra2 marker of stress granules. Measuring internal dynamics of membraneless organelles provides insight into their functional state, physical properties, and composition. Photoconversion has the advantage over other imaging techniques in that it is less phototoxic and allows for dual color tracking of proteins. For complete details on the use and execution of this protocol, please refer to Amen and Kaganovich (2020).<br />Competing Interests: The authors declare no competing interests.<br /> (© 2020 The Author(s).)
- Subjects :
- Benzothiazoles chemistry
Biomolecular Condensates metabolism
Biomolecular Condensates physiology
HEK293 Cells
Humans
Molecular Dynamics Simulation
Molecular Probes chemistry
Molecular Probes genetics
Organelles metabolism
Proteins metabolism
Stress Granules physiology
Molecular Probe Techniques instrumentation
Optical Imaging methods
Stress Granules metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2666-1667
- Volume :
- 1
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- STAR protocols
- Publication Type :
- Academic Journal
- Accession number :
- 33377110
- Full Text :
- https://doi.org/10.1016/j.xpro.2020.100217