In vivo protein trapping produces a functional expression codex of the vertebrate proteome
Genre
Journal ArticleDate
2011-06-01Author
Clark, KJBalciunas, D
Pogoda, HM
Ding, Y
Westcot, SE
Bedell, VM
Greenwood, TM
Urban, MD
Skuster, KJ
Petzold, AM
Ni, J
Nielsen, AL
Patowary, A
Scaria, V
Sivasubbu, S
Xu, X
Hammerschmidt, M
Ekker, SC
Subject
AnimalsAnimals, Genetically Modified
DNA Transposable Elements
Gene Expression Profiling
Gene Knockdown Techniques
Models, Animal
Molecular Sequence Data
Mutagenesis, Insertional
Proteome
Proteomics
Zebrafish
Zebrafish Proteins
Permanent link to this record
http://hdl.handle.net/20.500.12613/6038
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Show full item recordDOI
10.1038/nmeth.1606Abstract
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. © 2011 Nature America, Inc. All rights reserved.Citation to related work
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http://dx.doi.org/10.34944/dspace/6020