Format

Send to

Choose Destination
Nat Methods. 2009 Oct;6(10):745-51. doi: 10.1038/nmeth.1370. Epub 2009 Sep 6.

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

Author information

1
Max Delbrück Center for Molecular Medicine, Berlin, Germany.

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.

PMID:
19734907
PMCID:
PMC2756031
DOI:
10.1038/nmeth.1370
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center