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

Publication Type:

Journal Article

Source:

Nature methods, Volume 8, Issue 6, p.506-15 (2011)

Keywords:

Animalsdigestive disease, digestive deseases Animals, Genetically Modifieddigestive disease, digestive deseases DNA Transposable Elementsdigestive disease, digestive deseases Gene Expression Profilingdigestive disease, digestive deseases Gene Knockdown Techniquesdigestive disease, digestive deseases Models, Animaldigestive disease, digestive deseases Molecular Sequence Datadigestive disease, digestive deseases Mutagenesis, Insertionaldigestive disease, digestive deseases Proteomedigestive disease, digestive deseases Proteomicsdigestive disease, digestive deseases Zebrafishdigestive disease, digestive deseases Zebrafish Proteins

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.