title: Large-scale sorting of C. elegans embryos reveals the dynamics of small RNA expression.
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.
authors.affiliation: Max Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Marlon
authors.initials: M
authors.lastname: Stoeckius
authors.forename: Jonas
authors.initials: J
authors.lastname: Maaskola
authors.forename: Teresa
authors.initials: T
authors.lastname: Colombo
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Rahn
authors.forename: Marc R
authors.initials: MR
authors.lastname: Friedländer
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.forename: Fabio
authors.initials: F
authors.lastname: Piano
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Rajewsky
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000145:classification / Q000196:embryology* / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D034741:RNA, Small Interfering / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering