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| Status |
Public on Jul 17, 2012 |
| Title |
piRNAs initiate an epigenetic memory of non-self RNA in the C. elegans germline. |
| Organism |
Caenorhabditis elegans |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
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| Summary |
Organisms exhibit a fascinating array of gene-silencing pathways, which have evolved in part, to confront invasive nucleic acids such as transposons and viruses. A key question raised by the existence of these pathways is how do they distinguish “self” from “non-self” nucleic acids? Evidence exists for a number of mechanisms that might facilitate detection of foreign sequences including mechanisms that sense copy-number, unpaired DNA, or aberrant RNA (e.g.dsRNA). Here we describe an RNA-induced epigenetic silencing pathway, RNAe, that permanently silences single-copy transgenes. We show that the Piwi Argonaute PRG-1 and its genomically encoded piRNA cofactors initiate RNAe, while maintenance depends on chromatin factors and the WAGO Argonaute pathway. Our findings support a model in which PRG-1 scans for foreign sequences, while two other Argonaute pathways serve as epigenetic memories of "self" and "non-self" RNAs. These findings suggest how organisms may utilize RNAi-related mechanisms not only to recognize and silence foreign genes, but also to keep inventory of all genes expressed in the germ-line.
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| Overall design |
Examine small RNA population changes in different transgene lines. FLAG::WAGO-9 was immunoprecipitated from 20 mg of lysate essentially as described (Gu et al., 2009). Small RNAs were extracted from WAGO-9 immune complexes as well as a portion of the input lysate, gel-purified, pre-treated with TAP, cloned and sequenced as described (Gu et al., 2009).
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| Contributor(s) |
Shirayama M, Seth M, Lee H, Gu W, Ishidate T, Conte D, Mello C |
| Citation(s) |
22738726, 29456136 |
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| Submission date |
Jun 14, 2012 |
| Last update date |
Jul 26, 2019 |
| Contact name |
Craig Mello |
| Organization name |
University of Massachusetts Medical School
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| Department |
Program in Molecular Medicine
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| Lab |
Craig Mello
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| Street address |
368 Plantatoin Street, Suite AS5-2047
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| City |
Worcester |
| State/province |
MA |
| ZIP/Postal code |
01605 |
| Country |
USA |
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| Platforms (1) |
| GPL9269 |
Illumina Genome Analyzer II (Caenorhabditis elegans) |
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| Samples (11)
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| Relations |
| BioProject |
PRJNA168501 |
| SRA |
SRP013750 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE38724_RAW.tar |
89.7 Mb |
(http)(custom) |
TAR (of FA) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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