Monitoring condensate dynamics in S. cerevisiae using fluorescence recovery after photobleaching

Macy L. Sprunger; Meredith E. Jackrel

doi:10.1016/j.xpro.2022.101592

Monitoring condensate dynamics in S. cerevisiae using fluorescence recovery after photobleaching

Research output: Contribution to journal › Article › peer-review

Abstract

This protocol describes the use of fluorescence recovery after photobleaching (FRAP) to investigate the dynamics of Matrin-3 (MATR3) condensates in live budding yeast. We detail how to generate yeast strains containing MATR3 with an enhanced green fluorescent protein (eGFP) tag and induce MATR3-eGFP expression. We provide steps to prepare slides of immobilized yeast cells and perform FRAP imaging and data analysis. This protocol can be broadly applied to study condensate dynamics of a range of proteins in different model systems. For complete details on the use and execution of this protocol, please refer to Sprunger et al. (2022).

Original language	English
Pages (from-to)	101592
Number of pages	1
Journal	STAR Protocols
Volume	3
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2022.101592
State	Published - Sep 16 2022

Keywords

Biophysics
Cell Biology
Microscopy
Model Organisms
Protein Biochemistry

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10.1016/j.xpro.2022.101592

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title = "Monitoring condensate dynamics in S. cerevisiae using fluorescence recovery after photobleaching",

abstract = "This protocol describes the use of fluorescence recovery after photobleaching (FRAP) to investigate the dynamics of Matrin-3 (MATR3) condensates in live budding yeast. We detail how to generate yeast strains containing MATR3 with an enhanced green fluorescent protein (eGFP) tag and induce MATR3-eGFP expression. We provide steps to prepare slides of immobilized yeast cells and perform FRAP imaging and data analysis. This protocol can be broadly applied to study condensate dynamics of a range of proteins in different model systems. For complete details on the use and execution of this protocol, please refer to Sprunger et al. (2022).",

keywords = "Biophysics, Cell Biology, Microscopy, Model Organisms, Protein Biochemistry",

author = "Sprunger, {Macy L.} and Jackrel, {Meredith E.}",

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year = "2022",

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AU - Sprunger, Macy L.

AU - Jackrel, Meredith E.

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N2 - This protocol describes the use of fluorescence recovery after photobleaching (FRAP) to investigate the dynamics of Matrin-3 (MATR3) condensates in live budding yeast. We detail how to generate yeast strains containing MATR3 with an enhanced green fluorescent protein (eGFP) tag and induce MATR3-eGFP expression. We provide steps to prepare slides of immobilized yeast cells and perform FRAP imaging and data analysis. This protocol can be broadly applied to study condensate dynamics of a range of proteins in different model systems. For complete details on the use and execution of this protocol, please refer to Sprunger et al. (2022).

AB - This protocol describes the use of fluorescence recovery after photobleaching (FRAP) to investigate the dynamics of Matrin-3 (MATR3) condensates in live budding yeast. We detail how to generate yeast strains containing MATR3 with an enhanced green fluorescent protein (eGFP) tag and induce MATR3-eGFP expression. We provide steps to prepare slides of immobilized yeast cells and perform FRAP imaging and data analysis. This protocol can be broadly applied to study condensate dynamics of a range of proteins in different model systems. For complete details on the use and execution of this protocol, please refer to Sprunger et al. (2022).

KW - Biophysics

KW - Cell Biology

KW - Microscopy

KW - Model Organisms

KW - Protein Biochemistry

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AN - SCOPUS:85135424907

SN - 2666-1667

VL - 3

SP - 101592

JO - STAR Protocols

JF - STAR Protocols

IS - 3

ER -

Monitoring condensate dynamics in S. cerevisiae using fluorescence recovery after photobleaching

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Keywords

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