title: Evaluation of TRAP-sequencing technology with a versatile conditional mouse model.
abstract: Gene expression profiling of various cell lineages has provided invaluable insights into the molecular mechanisms regulating cellular development and differentiation. However, in vivo molecular profiling of rare and interspersed cell populations, such as endothelial cells, has remained challenging. We have generated a versatile floxed translating ribosome affinity purification (TRAP) mouse model, mCherryTRAP, for Cre-dependent translational profiling of distinct cell lineages from intact tissues. To identify cell type-specific transcripts using TRAP, the data have to be filtered to remove both background transcripts not expressed in the profiled cell type and transcripts expressed in all cell populations of the tissue/organ. Filtering has previously been achieved using transcribed RNA from the tissue/organ. Using the mCherryTRAP model, we demonstrate extensive differential expression of RNAs between the translatome and transcriptome of embryonic brains and kidneys. We evaluate the implications of these data for TRAP studies of abundant and rare cell populations. Finally, we demonstrate the applicability of the technology to study organ-specific endothelial cell differentiation.
authors.affiliation: Ludwig Institute for Cancer Research Ltd, Box 240, Stockholm SE-171 77, Sweden, Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm SE-171 77, Sweden and Max-Planck-Institute for Molecular Biomedicine, Department of Tissue Morphogenesis, and University of Münster, Faculty of Medicine, D-48149 Muenster, Germany.
authors.forename: Mike
authors.initials: M
authors.lastname: Hupe
authors.forename: Minerva Xueting
authors.initials: MX
authors.lastname: Li
authors.forename: Karin
authors.initials: K
authors.lastname: Gertow Gillner
authors.forename: Ralf H
authors.initials: RH
authors.lastname: Adams
authors.forename: Jan M
authors.initials: JM
authors.lastname: Stenman
mesh_terms: D000818:Animals; D001921:Brain / Q000196:embryology / Q000378:metabolism; D002454:Cell Differentiation; D042783:Endothelial Cells / Q000166:cytology; D020869:Gene Expression Profiling; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice / Q000235:genetics*; D023421:Models, Animal; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000032:analysis; D017423:Sequence Analysis, RNA; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein