A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila.
Elife
; 42015 Mar 31.
Article
en En
| MEDLINE
| ID: mdl-25824290
ABSTRACT
Here, we document a collection of â¼7434 MiMIC (Minos Mediated Integration Cassette) insertions of which 2854 are inserted in coding introns. They allowed us to create a library of 400 GFP-tagged genes. We show that 72% of internally tagged proteins are functional, and that more than 90% can be imaged in unfixed tissues. Moreover, the tagged mRNAs can be knocked down by RNAi against GFP (iGFPi), and the tagged proteins can be efficiently knocked down by deGradFP technology. The phenotypes associated with RNA and protein knockdown typically correspond to severe loss of function or null mutant phenotypes. Finally, we demonstrate reversible, spatial, and temporal knockdown of tagged proteins in larvae and adult flies. This new strategy and collection of strains allows unprecedented in vivo manipulations in flies for many genes. These strategies will likely extend to vertebrates.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Elementos Transponibles de ADN
/
Biblioteca de Genes
/
Mutagénesis Insercional
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Proteínas de Drosophila
/
Interferencia de ARN
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Drosophila melanogaster
Límite:
Animals
Idioma:
En
Revista:
Elife
Año:
2015
Tipo del documento:
Article
País de afiliación:
Estados Unidos