Translating ribosome affinity purification (TRAP) for cell-specific translation profiling in developing flowers.

The development of a multicellular organism is accompanied by cell differentiation. In fact, many biological processes have cell specificity, such that distinct cell types respond differently to endogenous or environmental cues. To obtain cell-specific gene expression profiles, translating ribosome affinity purification (TRAP) has been developed to label polysomes containing translating mRNAs in genetically defined cell types. Here, we describe the immunopurification of epitope-labeled polysomes and associated RNAs from target cell types. TRAP has the additional advantage of obtaining only translating mRNAs, which are a better proxy to the proteome than a standard mRNA preparation.

[1]  P. Greengard,et al.  A Translational Profiling Approach for the Molecular Characterization of CNS Cell Types , 2008, Cell.

[2]  Ying Wang,et al.  Advances in plant cell type-specific genome-wide studies of gene expression , 2011, Frontiers in Biology.

[3]  D. Shasha,et al.  A Gene Expression Map of the Arabidopsis Root , 2003, Science.

[4]  S. Henikoff,et al.  A simple method for gene expression and chromatin profiling of individual cell types within a tissue. , 2010, Developmental cell.

[5]  D. Galbraith,et al.  Global Characterization of Cell-Specific Gene Expression through Fluorescence-Activated Sorting of Nuclei1[W][OA] , 2008, Plant Physiology.

[6]  M. Arbeitman,et al.  A Versatile Method for Cell-Specific Profiling of Translated mRNAs in Drosophila , 2012, PloS one.

[7]  J. Bailey-Serres,et al.  Dynamic Light Regulation of Translation Status in Arabidopsis thaliana , 2012, Front. Plant Sci..

[8]  B. Mueller‐Roeber,et al.  Translatome and metabolome effects triggered by gibberellins during rosette growth in Arabidopsis , 2012, Journal of experimental botany.

[9]  E. Meyerowitz,et al.  Cell-type specific analysis of translating RNAs in developing flowers reveals new levels of control , 2010, Molecular Systems Biology.

[10]  I. Sussex,et al.  Laser Capture Microdissection of Cells from Plant Tissues1 , 2003, Plant Physiology.

[11]  J. Fletcher,et al.  Molecular mechanisms of flower development: an armchair guide , 2005, Nature Reviews Genetics.

[12]  R. Palmiter,et al.  Cell-type-specific isolation of ribosome-associated mRNA from complex tissues , 2009, Proceedings of the National Academy of Sciences.

[13]  P. Benfey,et al.  Cell type-specific transcriptional profiling: implications for metabolite profiling. , 2012, The Plant journal : for cell and molecular biology.

[14]  D. Galbraith,et al.  Profiling translatomes of discrete cell populations resolves altered cellular priorities during hypoxia in Arabidopsis , 2009, Proceedings of the National Academy of Sciences.