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

We describe a conditional in vivo protein trap mutagenesis system that reveals spatio-temporal protein expression dynamics and assesses gene function in the vertebrate Danio rerio. Integration of pGBT-RP2 (RP2), a gene-breaking transposon containing a protein trap, efficiently disrupts gene expressi...

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Detalles Bibliográficos
Autores principales: Clark, Karl J., Balciunas, Darius, Pogoda, Hans-Martin, Ding, Yonghe, Westcot, Stephanie E., Bedell, Victoria M., Greenwood, Tammy M., Urban, Mark D., Skuster, Kimberly J., Petzold, Andrew M., Ni, Jun, Nielsen, Aubrey, Patowary, Ashok, Scaria, Vinod, Sivasubbu, Sridhar, Xu, Xiaolei, Hammerschmidt, Matthias, Ekker, Stephen C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3306164/
https://www.ncbi.nlm.nih.gov/pubmed/21552255
http://dx.doi.org/10.1038/nmeth.1606
Descripción
Sumario:We describe a conditional in vivo protein trap mutagenesis system that reveals spatio-temporal protein expression dynamics and assesses gene function in the vertebrate Danio rerio. Integration of pGBT-RP2 (RP2), a gene-breaking transposon containing a protein trap, efficiently disrupts gene expression with >97% knockdown of normal transcript levels while simultaneously reporting protein expression of each locus. The mutant alleles are revertible in somatic tissues via Cre recombinase or splice-site blocking morpholinos, thus representing the first systematic conditional mutant alleles outside the mouse model. We report a collection of 350 zebrafish lines including a diverse array of 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 towards a unified codex of protein expression and direct functional annotation of the vertebrate genome.