Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Evidence of off-target effects associated with long dsRNAs in Drosophila cell-based assays

ABSTRACT: To evaluate the specificity of long dsRNAs used in high-throughput RNAi screens performed at the Drosophila RNAi Screening Center (DRSC), we performed a global analysis of their activity in 30 genome-wide screens completed at our facility. Surprisingly, our analysis predicts that dsRNAs containing ≥19 nucleotide perfect matches identified in silico to unintended targets may contribute to a significant false positive error rate arising from off-target effects. We confirmed experimentally that such sequences in dsRNAs lead to false positives and to the efficient knockdown of a cross-hybridizing transcript, raising a cautionary note when interpreting results based on the use of a single dsRNA per gene. Although a full appreciation of all causes of false positive errors remains to be determined, we suggest simple guidelines to help ensure high quality information from RNAi high-throughput screens Keywords: Specificity of long dsRNAs, Drosophila melanogaster, SL2 cell line, custom cDNA arrays, off-target effects, Drosophila RNAi Screening Center (DRSC)

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE5554 | GEO | 2006/08/18

SECONDARY ACCESSION(S): PRJNA96053

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Endocytosed dsRNAs induce lysosomal membrane permeabilization that allows cytosolic dsRNA translocation for Drosophila RNAi response

Project description:RNA interference (RNAi) is a gene-silencing mechanism triggered by the cytosolic entry of double-stranded RNAs (dsRNAs). Many animal cells internalize extracellular dsRNAs via endocytosis for RNAi induction. However, it is not clear how the endocytosed dsRNAs are translocated into the cytosol across the endo/lysosomal membrane. Herein, we show that in Drosophila S2 cells, endocytosed dsRNAs induce lysosomal membrane permeabilization (LMP) that allows cytosolic dsRNA translocation. LMP mediated by dsRNAs requires the lysosomal Cl−/H+ antiporter ClC-b/DmOstm1. In clc-b or dmostm1 knockout S2 cells, extracellular dsRNAs are endocytosed and reach the lysosomes normally but fail to enter the cytosol. Pharmacological induction of LMP restores extracellular dsRNA-directed RNAi in clc-b or dmostm1-knockout cells. Furthermore, clc-b or dmostm1 mutant flies are defective in extracellular dsRNA-directed RNAi and its associated antiviral immunity. Therefore, endocytosed dsRNAs have an intrinsic ability to induce ClC-b/DmOstm1-dependent LMP that allows cytosolic dsRNA translocation for RNAi responses in Drosophila cells.

2024-07-07 | GSE244523 | GEO

Endocytosed dsRNAs induce lysosomal membrane permeabilization that allows cytosolic dsRNA translocation for Drosophila RNAi response

Project description:Endocytosed dsRNAs induce lysosomal membrane permeabilization that allows cytosolic dsRNA translocation for Drosophila RNAi response

| PRJNA1023479 | ENA

modENCODE Drosophila RNA Binding Protein RNAi RNA-Seq Studies

Project description:Deep Sequencing of mRNA from the Drosophila melanogaster S2-DRSC cells that have been RNAi depleted of mRNAs encoding RNA binding proteins. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf

2009-10-09 | GSE18508 | GEO

PTGS induced by different dsRNAs

Project description:Unexpected variations in posttranscriptional silencing induced by differentially produced dsRNAs

| PRJEB33605 | ENA

modENCODE Drosophila RNA Binding Protein RNAi RNA-Seq Studies

2009-10-09 | E-GEOD-18508 | biostudies-arrayexpress

Effect of p53 on the expression of combo induced dsRNAs in 2153L GEM tumors

Project description:We analyzed the expression of dsRNAs in control and p53 restored 2153L GEM tumors upon different treatments. Our ChIP-seq and RNA-seq experiments showed that the restored p53 bound to the regulatory regions of dsRNA-containing genes that account for 91% of the total dsRNA abundance in combination therapy–treated 2153L tumors, and suppressed their expression upon combination therapy. The abundance of dsRNA (TPM) was calculated from the dsRIP experiment.

2024-10-16 | GSE276354 | GEO

modENCODE RNA-Seq of Drosophila cell lines Kc167, CME_W1_Cl.8+, S2-DRSC and ML-DmBG3-c2

Project description:Deep Sequencing of mRNA from the Drosophila melanogaster cell lines Kc167, CME_W1_Cl.8+, S2-DRSC and ML-DmBG3-c2. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf

2009-04-08 | GSE15596 | GEO

Gene expression profiling of S2 cells and S2 cells expressing AbdA

Project description:We sought to determine the genes regulated by the Drosophila Hox protein AbdA in a homogenous cell system. S2-DRSC cells that have no Hox expression were stably transfected with HA-tagged AbdA under the control of a metallothionein promoter.

2017-08-31 | GSE101555 | GEO

Identifying the targets of the pair-rule transcription factors Even-skipped and Runt in the late Drosophila blastoderm

Project description:It has long been appreciated that striped pair-rule transcription factor expression is necessary for convergent extension in the early Drosophila embryo, although the mechanisms that link these transcriptional regulators to planar polarity in this tissue have long been elusive. The goal of this study was to determine the transcriptional tragets of the pair-rule transcription factors Eve and Runt in Drosophila blastoderm embryos. We compared the transcriptional profiles of late blastoderm embryos injected with either water or dsRNAs against both eve and runt to identify differentially expressed genes that may directly contribute to the establishment of planar polarity during Drosophila convergent extension. Comparing the mRNA profiles from late blastoderm Drosophila embryos injected with either water (Water) or eve+runt dsRNAs (Eve), in triplicate, using Illumina HiSeq.

2014-11-02 | E-GEOD-61689 | biostudies-arrayexpress

Production of functional small interfering RNAs by human Dicer

Project description:While RNA interference (RNAi) functions as a potent antiviral innate immune response in plants and invertebrates, mammalian somatic cells appear incapable of mounting an RNAi response and few small interfering RNAs (siRNAs) can be detected. To examine why siRNA production is inefficient, we have generated double knockout human cells lacking both Dicer and PKR. Using these cells, which tolerate dsRNA expression, we show that mutant forms of human Dicer lacking the amino-terminal helicase domain can process dsRNAs to produce high levels of siRNAs that are readily detectable by Northern blot and that can effectively and specifically inhibit the expression of cognate mRNAs. However, even these more active Dicer mutants produce only modest levels of viral siRNAs in infected cells that are insufficient to inhibit viral replication. We conclude that the production of siRNAs from viral dsRNAs is likely inefficient due to the poor accessibility of viral dsRNAs to Dicer.

2015-12-02 | GSE69433 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data