In vivo protein trapping produces a functional expression codex of the vertebrate proteome.
Nat Methods
; 8(6): 506-15, 2011 Jun.
Article
em En
| MEDLINE
| ID: mdl-21552255
ABSTRACT
We describe a conditional in vivo protein-trap mutagenesis system that reveals spatiotemporal protein expression dynamics and can be used to assess gene function in the vertebrate Danio rerio. Integration of pGBT-RP2.1 (RP2), a gene-breaking transposon containing a protein trap, efficiently disrupts gene expression with >97% knockdown of normal transcript amounts and simultaneously reports protein expression for each locus. The mutant alleles are revertible in somatic tissues via Cre recombinase or splice-site-blocking morpholinos and are thus to our knowledge the first systematic conditional mutant alleles outside the mouse model. We report a collection of 350 zebrafish lines that include diverse molecular loci. RP2 integrations reveal the complexity of genomic architecture and gene function in a living organism and can provide information on protein subcellular localization. The RP2 mutagenesis system is a step toward a unified 'codex' of protein expression and direct functional annotation of the vertebrate genome.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Peixe-Zebra
/
Mutagênese Insercional
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Proteoma
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Proteínas de Peixe-Zebra
Limite:
Animals
Idioma:
En
Revista:
Nat Methods
Assunto da revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Estados Unidos