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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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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
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author 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.
author_facet 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.
author_sort Clark, Karl J.
collection PubMed
description 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.
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spelling pubmed-33061642012-03-16 in vivo protein trapping produces a functional expression codex of the vertebrate proteome 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. Nat Methods Article 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. 2011-05-08 2011-06 /pmc/articles/PMC3306164/ /pubmed/21552255 http://dx.doi.org/10.1038/nmeth.1606 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
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.
in vivo protein trapping produces a functional expression codex of the vertebrate proteome
title in vivo protein trapping produces a functional expression codex of the vertebrate proteome
title_full in vivo protein trapping produces a functional expression codex of the vertebrate proteome
title_fullStr in vivo protein trapping produces a functional expression codex of the vertebrate proteome
title_full_unstemmed in vivo protein trapping produces a functional expression codex of the vertebrate proteome
title_short in vivo protein trapping produces a functional expression codex of the vertebrate proteome
title_sort in vivo protein trapping produces a functional expression codex of the vertebrate proteome
topic Article
url 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
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