Isolation, Profiling, and Tracking of Extracellular Vesicle Cargo in Caenorhabditis Elegans

Overview

Journal Curr Biol

Publisher Cell Press

Specialty Biology

Date 2022 Mar 25

PMID 35334227

Authors

Inna A Nikonorova

Juan Wang

Alexander L Cope

Peter E Tilton

Kaiden M Power

Jonathon D Walsh

Jyothi S Akella

Amber R Krauchunas

Premal Shah

Maureen M Barr

Affiliations

Soon will be listed here.

Abstract

Extracellular vesicles (EVs) may mediate intercellular communication by carrying protein and RNA cargo. The composition, biology, and roles of EVs in physiology and pathology have been primarily studied in the context of biofluids and in cultured mammalian cells. The experimental tractability of C. elegans makes for a powerful in vivo animal system to identify and study EV cargo from its cellular source. We developed an innovative method to label, track, and profile EVs using genetically encoded, fluorescent-tagged EV cargo and conducted a large-scale isolation and proteomic profiling. Nucleic acid binding proteins (∼200) are overrepresented in our dataset. By integrating our EV proteomic dataset with single-cell transcriptomic data, we identified and validated ciliary EV cargo: CD9-like tetraspanin (TSP-6), ectonucleotide pyrophosphatase/phosphodiesterase (ENPP-1), minichromosome maintenance protein (MCM-3), and double-stranded RNA transporter SID-2. C. elegans EVs also harbor RNA, suggesting that EVs may play a role in extracellular RNA-based communication.

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