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E-MTAB-8067

ABSTRACT: RNA-seq of CK2alpha-silenced H9-c2 derived myoblasts using siRNA or doxycyclin inducible shRNA.

PROVIDER: E-MTAB-8067 | ExpressionAtlas | 2019-08-30

REPOSITORIES: ExpressionAtlas

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			Action	DRS
	E-MTAB-8067-analysis-methods.tsv	Tabular
	E-MTAB-8067-analytics.tsv	Tabular
	E-MTAB-8067-atlasExperimentSummary.Rdata	Rdata
	E-MTAB-8067-configuration.xml	Xml
	E-MTAB-8067-g2_g1-mvaPlot.png	Other

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RNA-seq of CK2alpha-silenced H9-c2 derived myoblasts using siRNA or doxycyclin inducible shRNA.

Project description:RNA-seq of CK2alpha-silenced H9-c2 derived myoblasts using siRNA or doxycyclin inducible shRNA.

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Widespread siRNA "off-target" transcript silencing mediated by seed region sequence complementarity

Project description:Transfected siRNAs and miRNAs regulate numerous transcripts that have only limited complementarity to the active strand of the RNA duplex. This process reflects natural target regulation by miRNAs, but is an unintended (“off-target”) consequence of siRNA-mediated silencing. Here we demonstrate that this unintended off-target silencing is widespread, and occurs in a manner reminiscent of target silencing by miRNAs. A high proportion of unintended transcripts silenced by siRNAs showed 3’ UTR sequence complementarity to the seed region of the siRNA. Base mismatches within the siRNA seed region reduced the set of original off-target transcripts but generated new sets of silenced transcripts with sequence complementarity to the mismatched seed sequence. An inducible shRNA silenced a subset of transcripts that were silenced by an siRNA of the same sequence, demonstrating that unintended silencing is sequence-mediated and is independent of delivery method. In all cases, off-target transcript silencing was accompanied by loss of the corresponding protein and occurred with similar dependence on siRNA concentration as silencing of the target transcript. These results demonstrate that short stretches of sequence complementarity to the seed region of the siRNA are key to the silencing of unintended transcripts, and that this limits the specificity of siRNA-mediated transcript silencing. Because these off-target events are sequence-dependent, inclusion of multiple independent siRNAs to the target of interest can help to distinguish true positives from false positives in functional genetic analyses. Keywords: siRNA, RNAi, sequence alignment, off-target, seed region

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2019-12-17 | PXD016535 | Pride

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