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Nat Methods. 2011 Jun;8(6):506-15. doi: 10.1038/nmeth.1606. Epub 2011 May 8.

In vivo protein trapping produces a functional expression codex of the vertebrate proteome.

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1
Mayo Clinic, Rochester, Minnesota, USA.

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.

PMID:
21552255
PMCID:
PMC3306164
DOI:
10.1038/nmeth.1606
[Indexed for MEDLINE]
Free PMC Article
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