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Thomas, A., Lee, P.J., Dalton, J.E., Nomie, K.J., Stoica, L., Costa-Mattioli, M., Chang, P., Nuzhdin, S., Arbeitman, M.N., Dierick, H.A. (2012). A Versatile Method for Cell-Specific Profiling of Translated mRNAs in Drosophila.  PLoS ONE 7(7): e40276.
FlyBase ID
FBrf0218903
Publication Type
Research paper
Abstract

In Drosophila melanogaster few methods exist to perform rapid cell-type or tissue-specific expression profiling. A translating ribosome affinity purification (TRAP) method to profile actively translated mRNAs has been developed for use in a number of multicellular organisms although it has only been implemented to examine limited sets of cell- or tissue-types in these organisms. We have adapted the TRAP method for use in the versatile GAL4/UAS system of Drosophila allowing profiling of almost any tissue/cell-type with a single genetic cross. We created transgenic strains expressing a GFP-tagged ribosomal protein, RpL10A, under the control of the UAS promoter to perform cell-type specific translatome profiling. The GFP::RpL10A fusion protein incorporates efficiently into ribosomes and polysomes. Polysome affinity purification strongly enriches mRNAs from expected genes in the targeted tissues with sufficient sensitivity to analyze expression in small cell populations. This method can be used to determine the unique translatome profiles in different cell-types under varied physiological, pharmacological and pathological conditions.

PubMed ID
PubMed Central ID
PMC3391276 (PMC) (EuropePMC)
Related Publication(s)
Erratum

Correction: A Versatile Method for Cell-Specific Profiling of Translated mRNAs in Drosophila.
Thomas et al., 2013, PLoS ONE 8(7): [FBrf0222759]

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Secondary IDs
    Language of Publication
    English
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Journal
    Abbreviation
    PLoS ONE
    Title
    PLoS ONE
    Publication Year
    2006-
    ISBN/ISSN
    1932-6203
    Data From Reference
    Alleles (3)
    Genes (26)
    Natural transposons (1)
    Insertions (1)
    Experimental Tools (2)
    Transgenic Constructs (2)