In situ detection of ribonucleoprotein complex assembly in the C. Elegans germline using proximity ligation assay

Nicholas J. Day; Xiaobo Wang; Ekaterina Voronina

doi:10.3791/60982

In situ detection of ribonucleoprotein complex assembly in the C. Elegans germline using proximity ligation assay

Nicholas J. Day
, Xiaobo Wang
, Ekaterina Voronina

Division of Biological and Biomedical Sciences

University of Montana

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

8 Scopus citations

Abstract

Understanding when and where protein-protein interactions (PPIs) occur is critical to understanding protein function in the cell and how broader processes such as development are affected. The Caenorhabditis elegans germline is a great model system for studying PPIs that are related to the regulation of stem cells, meiosis, and development. There are a variety of well-developed techniques that allow proteins of interest to be tagged for recognition by standard antibodies, making this system advantageous for proximity ligation assay (PLA) reactions. As a result, the PLA is able to show where PPIs occur in a spatial and temporal manner in germlines more effectively than alternative approaches. Described here is a protocol for the application and quantification of this technology to probe PPIs in the C. elegans germline.

Original language	English
Article number	e60982
Journal	Journal of Visualized Experiments
Volume	2020
Issue number	159
DOIs	https://doi.org/10.3791/60982
State	Published - May 2020

Funding

Some nematode strains used in this study were provided by the Caenorhabditis Genetics Center funded by the NIH (P40OD010440). Confocal microscopy was performed in the University of Montana BioSpectroscopy Core Research Laboratory operated with support from NIH Awards P20GM103546 and S10OD021806. This work was supported in part by the NIH grant GM109053 to E.V., American Heart Association Fellowship 18PRE34070028 to X.W., and Montana Academy of Sciences award to X.W. The funders were not involved in study design or writing the report. We thank M. Ellenbecker for discussion.

Funders	Funder number
GM109053, P20GM103546, P40OD010440
S10OD021806
American Heart Association	18PRE34070028

Keywords

C. Elegans
Developmental Biology
Germline
Issue 159
LC8
PLA
Proximity ligation assay
RNA-binding protein

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10.3791/60982

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title = "In situ detection of ribonucleoprotein complex assembly in the C. Elegans germline using proximity ligation assay",

abstract = "Understanding when and where protein-protein interactions (PPIs) occur is critical to understanding protein function in the cell and how broader processes such as development are affected. The Caenorhabditis elegans germline is a great model system for studying PPIs that are related to the regulation of stem cells, meiosis, and development. There are a variety of well-developed techniques that allow proteins of interest to be tagged for recognition by standard antibodies, making this system advantageous for proximity ligation assay (PLA) reactions. As a result, the PLA is able to show where PPIs occur in a spatial and temporal manner in germlines more effectively than alternative approaches. Described here is a protocol for the application and quantification of this technology to probe PPIs in the C. elegans germline.",

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AB - Understanding when and where protein-protein interactions (PPIs) occur is critical to understanding protein function in the cell and how broader processes such as development are affected. The Caenorhabditis elegans germline is a great model system for studying PPIs that are related to the regulation of stem cells, meiosis, and development. There are a variety of well-developed techniques that allow proteins of interest to be tagged for recognition by standard antibodies, making this system advantageous for proximity ligation assay (PLA) reactions. As a result, the PLA is able to show where PPIs occur in a spatial and temporal manner in germlines more effectively than alternative approaches. Described here is a protocol for the application and quantification of this technology to probe PPIs in the C. elegans germline.

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KW - LC8

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KW - RNA-binding protein

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In situ detection of ribonucleoprotein complex assembly in the C. Elegans germline using proximity ligation assay

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