Rapid Molecular Profiling of Defined Cell Types Using Viral TRAP.

Abstract

Translational profiling methodologies enable the systematic characterization of cell types in complex tissues, such as the mammalian brain, where neuronal isolation is exceptionally difficult. Here, we report a versatile strategy for profiling CNS cell types in a spatiotemporally restricted fashion by engineering a Cre-dependent adeno-associated virus expressing an EGFP-tagged ribosomal protein (AAV-FLEX-EGFPL10a) to access translating mRNAs by translating ribosome affinity purification (TRAP). We demonstrate the utility of this AAV to target a variety of genetically and anatomically defined neural populations expressing Cre recombinase and illustrate the ability of this viral TRAP (vTRAP) approach to recapitulate the molecular profiles obtained by bacTRAP in corticothalamic neurons across multiple serotypes. Furthermore, spatially restricting adeno-associated virus (AAV) injections enabled the elucidation of regional differences in gene expression within this cell type. Altogether, these results establish the broad applicability of the vTRAP strategy for the molecular dissection of any CNS or peripheral cell type that can be engineered to express Cre.

DOI: 10.1016/j.celrep.2017.03.048

Cite this paper

@article{Nectow2017RapidMP, title={Rapid Molecular Profiling of Defined Cell Types Using Viral TRAP.}, author={Alexander R. Nectow and Maria V Moya and Mats I. Ekstrand and Awni Mousa and Kelly L McGuire and Caroline Sferrazza and Bianca Field and Gabrielle S Rabinowitz and Kirsty Sawicka and Yupu Liang and Jeffrey M. Friedman and Nathaniel Heintz and Eric F Schmidt}, journal={Cell reports}, year={2017}, volume={19 3}, pages={655-667} }