4.8 Article

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

Journal

NATURE METHODS
Volume 6, Issue 10, Pages 745-U16

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1370

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Funding

  1. NHGRI NIH HHS [U01 HG004276, U01 HG004276-03] Funding Source: Medline
  2. NICHD NIH HHS [R01HD046236, R01 HD046236, R01 HD046236-06] Funding Source: Medline

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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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