Evidence of off-target effects associated with long dsRNAs in Drosophila melanogaster cell-based assays.
Nat Methods
; 3(10): 833-8, 2006 Oct.
Article
en En
| MEDLINE
| ID: mdl-16964256
ABSTRACT
To evaluate the specificity of long dsRNAs used in high-throughput RNA interference (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. Notably, our analysis predicts that dsRNAs containing > or = 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 efficient knockdown of a cross-hybridizing transcript, raising a cautionary note about 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.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
ARN Bicatenario
/
Pruebas Genéticas
/
Interferencia de ARN
/
Drosophila melanogaster
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
/
Risk_factors_studies
Límite:
Animals
Idioma:
En
Revista:
Nat Methods
Asunto de la revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Año:
2006
Tipo del documento:
Article
País de afiliación:
Estados Unidos