A comprehensive survey of C. elegans argonaute proteins reveals organism-wide gene regulatory networks and functions.

A comprehensive survey of C. elegans argonaute proteins reveals organism-wide gene regulatory networks and functions.

eLife (2023-02-16)

Uri Seroussi, Andrew Lugowski, Lina Wadi, Robert X Lao, Alexandra R Willis, Winnie Zhao, Adam E Sundby, Amanda G Charlesworth, Aaron W Reinke, Julie M Claycomb

PMID36790166

ABSTRACT

Argonaute (AGO) proteins associate with small RNAs to direct their effector function on complementary transcripts. The nematode Caenorhabditis elegans contains an expanded family of 19 functional AGO proteins, many of which have not been fully characterized. In this work, we systematically analyzed every C. elegans AGO using CRISPR-Cas9 genome editing to introduce GFP::3xFLAG tags. We have characterized the expression patterns of each AGO throughout development, identified small RNA binding complements, and determined the effects of ago loss on small RNA populations and developmental phenotypes. Our analysis indicates stratification of subsets of AGOs into distinct regulatory modules, and integration of our data led us to uncover novel stress-induced fertility and pathogen response phenotypes due to ago loss.

MATERIALS

Product Number

Brand

Product Description

F1804

Sigma-Aldrich

Monoclonal ANTI-FLAG^® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)

P5726

Sigma-Aldrich

Phosphatase Inhibitor Cocktail 2, aqueous solution (dark coloration may develop upon storage, which does not affect the activity)

P0044

Sigma-Aldrich

Phosphatase Inhibitor Cocktail 3, DMSO solution

11836170001

Roche

cOmplete^™, Mini, EDTA-free Protease Inhibitor Cocktail, Protease Inhibitor Cocktail Tablets provided in a glass vial, Tablets provided in a glass vial