ZFIN ID: ZDB-PUB-111117-23
A versatile gene trap to visualize and interrogate the function of the vertebrate proteome
Trinh, A., Hochgreb, T., Graham, M., Wu, D., Ruf-Zamojski, F., Jayasena, C.S., Saxena, A., Hawk, R., Gonzalez-Serricchio, A., Dixson, A., Chow, E., Gonzales, C., Leung, H.Y., Solomon, I., Bronner-Fraser, M., Megason, S.G., and Fraser, S.E.
Date: 2011
Source: Genes & Development   25(21): 2306-20 (Journal)
Registered Authors: Bronner-Fraser, Marianne, Chow, Elly, Dixson, Alana, Fraser, Scott E., Gonzalez-Serricchio, Aidyl, Hawk, Rasheeda, Hochgreb, Tatiana, Jayasena, Chaturani (Saku), Megason, Sean, Ruf-Zamojski, Frederique, Saxena, Ankur, Trinh, Le
Keywords: endogenous proteins, proteome, conditional mutagenesis, gene trap, genetic manipulation, gene knockdown
MeSH Terms:
  • Animals
  • Bacterial Proteins/genetics
  • Databases, Protein
  • Embryo, Nonmammalian
  • Genetic Vectors
  • Internet
  • Luminescent Proteins/genetics
  • Molecular Sequence Annotation
  • Mutation
  • Proteome*
  • Proteomics/methods*
  • Recombinant Fusion Proteins/genetics
  • Recombinant Fusion Proteins/metabolism
  • Zebrafish
PubMed: 22056673 Full text @ Genes & Dev.
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ABSTRACT
We report a multifunctional gene-trapping approach, which generates full-length Citrine fusions with endogenous proteins and conditional mutants from a single integration event of the FlipTrap vector. We identified 170 FlipTrap zebrafish lines with diverse tissue-specific expression patterns and distinct subcellular localizations of fusion proteins generated by the integration of an internal citrine exon. Cre-mediated conditional mutagenesis is enabled by heterotypic lox sites that delete Citrine and “flip” in its place mCherry with a polyadenylation signal, resulting in a truncated fusion protein. Inducing recombination with Cerulean-Cre results in fusion proteins that often mislocalize, exhibit mutant phenotypes, and dramatically knock down wild-type transcript levels. FRT sites in the vector enable targeted genetic manipulation of the trapped loci in the presence of Flp recombinase. Thus, the FlipTrap captures the functional proteome, enabling the visualization of full-length fluorescent fusion proteins and interrogation of function by conditional mutagenesis and targeted genetic manipulation.
ADDITIONAL INFORMATION