Large-scale Sorting of C. Elegans Embryos Reveals the Dynamics of Small RNA Expression

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2009 Sep 8

PMID 19734907

Citations 55

Authors

Marlon Stoeckius

Jonas Maaskola

Teresa Colombo

Hans-Peter Rahn

Marc R Friedlander

Na Li

Wei Chen

Fabio Piano

Nikolaus Rajewsky

Affiliations

Soon will be listed here.

Abstract

Caenorhabditis elegans is one of the most prominent model systems for embryogenesis, but collecting many precisely staged embryos has been impractical. Thus, early C. elegans embryogenesis has not been amenable to most high-throughput genomics or biochemistry assays. To overcome this problem, we devised a method to collect staged C. elegans embryos by fluorescence-activated cell sorting (eFACS). In a proof-of-principle experiment, we found that a single eFACS run routinely yielded tens of thousands of almost perfectly staged 1-cell stage embryos. As the earliest embryonic events are driven by posttranscriptional regulation, we combined eFACS with second-generation sequencing to profile the embryonic expression of small, noncoding RNAs. We discovered complex and orchestrated changes in the expression between and within almost all classes of small RNAs, including microRNAs and 26G-RNAs, during embryogenesis.

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