A resource for large-scale RNA-interference-based screens in mammals.

Paddison, Patrick J; Silva, Jose M; Conklin, Douglas S; Schlabach, Mike; Li, Mamie; Aruleba, Shola; Balija, Vivekanand; O'Shaughnessy, Andy; Gnoj, Lidia; Scobie, Kim; Chang, Kenneth; Westbrook, Thomas; Cleary, Michele; Sachidanandam, Ravi; McCombie, W Richard; Elledge, Stephen J; Hannon, Gregory J

Paddison, Patrick J; Silva, Jose M; Conklin, Douglas S; Schlabach, Mike; Li, Mamie; Aruleba, Shola; Balija, Vivekanand; O'Shaughnessy, Andy; Gnoj, Lidia; Scobie, Kim; Chang, Kenneth; Westbrook, Thomas; Cleary, Michele; Sachidanandam, Ravi; McCombie, W Richard; Elledge, Stephen J; Hannon, Gregory J.

Afiliación

Paddison PJ; Cold Spring Harbor Laboratory, Watson School of Biological Sciences, 1 Bungtown Road, Cold Spring Harbor, New York 11724, USA.

Nature ; 428(6981): 427-31, 2004 Mar 25.

Article en En | MEDLINE | ID: mdl-15042091

RESUMEN

Gene silencing by RNA interference (RNAi) in mammalian cells using small interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) has become a valuable genetic tool. Here, we report the construction and application of a shRNA expression library targeting 9,610 human and 5,563 mouse genes. This library is presently composed of about 28,000 sequence-verified shRNA expression cassettes contained within multi-functional vectors, which permit shRNA cassettes to be packaged in retroviruses, tracked in mixed cell populations by means of DNA 'bar codes', and shuttled to customized vectors by bacterial mating. In order to validate the library, we used a genetic screen designed to report defects in human proteasome function. Our results suggest that our large-scale RNAi library can be used in specific, genetic applications in mammals, and will become a valuable resource for gene analysis and discovery.

Asunto(s)

Biblioteca de Genes; Ingeniería Genética/métodos; Interferencia de ARN; ARN Interferente Pequeño/genética; Animales; Clonación Molecular; Cisteína Endopeptidasas/genética; Cisteína Endopeptidasas/metabolismo; Genes/genética; Vectores Genéticos; Humanos; Ratones; Complejos Multienzimáticos/genética; Complejos Multienzimáticos/metabolismo; Análisis de Secuencia por Matrices de Oligonucleótidos; Complejo de la Endopetidasa Proteasomal; ARN Interferente Pequeño/metabolismo; Reproducibilidad de los Resultados; Especificidad por Sustrato

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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ingeniería Genética / Biblioteca de Genes / ARN Interferente Pequeño / Interferencia de ARN Límite: Animals / Humans Idioma: En Revista: Nature Año: 2004 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido

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