Rapid Molecular Profiling of Defined Cell Types Using Viral TRAP

Publication date: 18 April 2017
Source:Cell Reports, Volume 19, Issue 3
Author(s): Alexander R. Nectow, Maria V. Moya, Mats I. Ekstrand, Awni Mousa, Kelly L. McGuire, Caroline E. Sferrazza, Bianca C. Field, Gabrielle S. Rabinowitz, Kirsty Sawicka, Yupu Liang, Jeffrey M. Friedman, Nathaniel Heintz, Eric F. Schmidt
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.

Graphical abstract

Teaser

Nectow et al. describe vTRAP, a technology to purify translating mRNAs from genetically defined cell types in a spatiotemporally restricted fashion. Multiplexing vTRAP with other technologies offers a comprehensive strategy to interrogate the precise role of individual, cell-type-specific genes in neural circuit function.


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