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Large-scale sorting of C. elegans embryos reveals the dynamics of small RNA expression

Official URL:https://doi.org/10.1038/nmeth.1370
PubMed:View item in PubMed
Creators Name:Stoeckius, M. and Maaskola, J. and Colombo, T. and Rahn, H.P. and Friedlaender, M.R. and Li, N. and Chen, W. and Piano, F. and Rajewsky, N.
Journal Title:Nature Methods
Journal Abbreviation:Nat Methods
Volume:6
Number:10
Page Range:745-751
Date:October 2009
Keywords:Nonmammalian Embryo, Flow Cytometry, Small Interfering RNA, Animals, Caenorhabditis Elegans
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.
ISSN:1548-7091
Publisher:Nature Publishing Group (U.S.A.)
Item Type:Article

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