Tissue-Specific Split sfGFP System for Streamlined Expression of GFP Tagged Proteins in the Caenorhabditis elegans Germline

Adam Hefel; Sarit Smolikove

doi:10.1534/g3.119.400162

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Tissue-Specific Split sfGFP System for Streamlined Expression of GFP Tagged Proteins in the Caenorhabditis elegans Germline

Journal article

Open access

Tissue-Specific Split sfGFP System for Streamlined Expression of GFP Tagged Proteins in the Caenorhabditis elegans Germline

Adam Hefel and Sarit Smolikove

G3 (Bethesda, Md.), Vol.9(6), pp.1933-1943

06/05/2019

DOI: 10.1534/g3.119.400162

PMCID: PMC6553534

PMID: 30992318

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https://doi.org/10.1534/g3.119.400162View

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Abstract

Identifying protein localization is a useful tool in analyzing protein function. Using GFP-fusion tags, researchers can study the function of endogenous proteins in living tissue. However, these tags are considerably large, making them difficult to insert, and they can potentially affect the normal function of these proteins. To improve on these drawbacks, we have adopted the split sfGFP system for studying the localization of proteins in the germline. This system divides the "super folder" GFP into 2 fragments, allowing researchers to use CRISPR/Cas9 to tag proteins more easily with the smaller subunit, while constitutively expressing the larger subunit from another locus. These two parts are able to stably interact, producing a functional GFP when both fragments are in the same cellular compartment. Our data demonstrate that the split sfGFP system can be adapted for use in to tag endogenous proteins with relative ease. Strains containing the tags are homozygous viable and fertile. These small subunit tags produce fluorescent signals that matched the localization patterns of the wild-type protein in the gonad. Thus, our study shows that this approach could be used for tissue-specific GFP expression from an endogenous locus.

Animals

Biomarkers

Caenorhabditis elegans - genetics

Caenorhabditis elegans - metabolism

Caenorhabditis elegans Proteins - genetics

Caenorhabditis elegans Proteins - metabolism

Cloning, Molecular

Fertility

Gene Expression

Germ Cells - metabolism

Green Fluorescent Proteins - genetics

Green Fluorescent Proteins - metabolism

Molecular Imaging

Recombinant Fusion Proteins - genetics

Recombinant Fusion Proteins - metabolism

Details

Title: Subtitle: Tissue-Specific Split sfGFP System for Streamlined Expression of GFP Tagged Proteins in the Caenorhabditis elegans Germline
Creators: Adam Hefel - Department of Biology, University of Iowa, Iowa City, IA 52240
Sarit Smolikove - Department of Biology, University of Iowa, Iowa City, IA 52240 sarit-smolikove@uiowa.edu
Resource Type: Journal article
Publication Details: G3 (Bethesda, Md.), Vol.9(6), pp.1933-1943
DOI: 10.1534/g3.119.400162
PMID: 30992318
PMCID: PMC6553534
NLM abbreviation: G3 (Bethesda)
ISSN: 2160-1836
eISSN: 2160-1836
Grant note: R01 GM112657 / NIGMS NIH HHS
Language: English
Date published: 06/05/2019
Academic Unit: Biology
Record Identifier: 9984217419702771

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