Large-scale sorting of C. elegans embryos reveals the dynamics of small RNA expression
Fluorescence-activated cell sorting of worm embryos promises to replace manual sorting of staged embryos and yields large populations highly enriched in specific developmental stages, allowing high-throughput genomic analysis. Caenorhabditis elegans is one of the most prominent model systems for emb...
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Veröffentlicht in: | Nature methods 2009-10, Vol.6 (10), p.745-751 |
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Sprache: | eng |
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Zusammenfassung: | Fluorescence-activated cell sorting of worm embryos promises to replace manual sorting of staged embryos and yields large populations highly enriched in specific developmental stages, allowing high-throughput genomic analysis.
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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ISSN: | 1548-7091 1548-7105 1548-7105 |
DOI: | 10.1038/nmeth.1370 |