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
-
- Animals
- Animals, Genetically Modified
- DNA Transposable Elements/genetics
- Gene Expression Profiling
- Gene Knockdown Techniques/methods
- Models, Animal
- Molecular Sequence Data
- Mutagenesis, Insertional/methods*
- Proteome/genetics*
- Proteomics/methods
- Zebrafish/genetics*
- Zebrafish Proteins/genetics*
- PubMed
- 21552255 Full text @ Nat. Methods
Citation
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. (2011) In vivo protein trapping produces a functional expression codex of the vertebrate proteome. Nature Methods. 8(6):506-512.
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