PUBLICATION
In vivo protein trapping produces a functional expression codex of the vertebrate proteome
- Authors
- Clark, K.J., Balciunas, D., Pogoda, H.M., Ding, Y., Westcot, S.E., Bedell, V.M., Greenwood, T.M., Urban, M.D., Skuster, K.J., Petzold, A.M., Ni, J., Nielsen, A.L., Patowary, A., Scaria, V., Sivasubbu, S., Xu, X., Hammerschmidt, M., and Ekker, S.C.
- ID
- ZDB-PUB-110523-11
- Date
- 2011
- Source
- Nature Methods 8(6): 506-512 (Journal)
- Registered Authors
- Balciunas, Darius, Bedell, Victoria, Clark, Karl, Ding, Yonghe, Ekker, Stephen C., Hammerschmidt, Matthias, Petzold, Andrew, Pogoda, Hans-Martin, Sivasubbu, Sridhar, Westcot, Stephanie, Xu, Xiaolei
- Keywords
- none
- MeSH Terms
-
- Proteome/genetics*
- Models, Animal
- Animals, Genetically Modified
- Molecular Sequence Data
- Zebrafish/genetics*
- PubMed
- 21552255 Full text @ Nat. Methods
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping