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Visualizing nuclear RNAi activity in single living human cells.

ABSTRACT: Nuclear RNA interference (RNAi) is mediated by the canonical RNAi machinery and can lead to transcriptional silencing, transcriptional activation, or modulation of alternative splicing patterns. These effects transpire through changes in histone and DNA modifications via RNAi-mediated recruitment of chromatin-modifying enzymes. To prove that nuclear RNAi occurs and modulates transcription in human cells, we used live-cell imaging to detect and track nuclear RNAi transcriptional repression in single living human cells. While employing reporter genes constructed with inducible promoters and cognate-inducible short hairpin RNA (shRNA) targeted against the reporter coding region, we have characterized the dynamics of the nuclear RNAi process in living human cells. We show that the silencing effect is mediated through the nascent mRNA, followed by activity of histone methylating enzymes, but not through DNA methylation.

SUBMITTER: Avivi S

PROVIDER: S-EPMC5651755 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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Visualizing nuclear RNAi activity in single living human cells.

Avivi Shira S Mor Amir A Dotan Iris I Tzadok Sivan S Kanter Itamar I Kinor Noa N Canaani Dan D Shav-Tal Yaron Y

Proceedings of the National Academy of Sciences of the United States of America 20171003 42

Nuclear RNA interference (RNAi) is mediated by the canonical RNAi machinery and can lead to transcriptional silencing, transcriptional activation, or modulation of alternative splicing patterns. These effects transpire through changes in histone and DNA modifications via RNAi-mediated recruitment of chromatin-modifying enzymes. To prove that nuclear RNAi occurs and modulates transcription in human cells, we used live-cell imaging to detect and track nuclear RNAi transcriptional repression in sin ...[more]

PMID: 29073029

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