Cell type–specific mRNA purification by translating ribosome affinity purification (TRAP)
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Heiman_Cell type-specific.pdf
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1.56 MB
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Author(s)
Heiman, Myriam
Kulicke, Ruth
Greengard, Paul
Heintz, Nathaniel
Fenster, Robert
Date Issued
May 2014
Journal
Nature Protocols
Publisher
Nature Publishing Group
Citation
Heiman, Myriam, Ruth Kulicke, Robert J Fenster, Paul Greengard, and Nathaniel Heintz. “Cell Type–specific mRNA Purification by Translating Ribosome Affinity Purification (TRAP).” Nature Protocols 9, no. 6 (May 8, 2014): 1282–1291.
Version
Author's final manuscript
Abstract
Cellular diversity and architectural complexity create barriers to understanding the function of the mammalian CNS at a molecular level. To address this problem, we have recently developed a methodology that provides the ability to profile the entire translated mRNA complement of any genetically defined cell population. This methodology, which we termed translating ribosome affinity purification, or TRAP, combines cell type–specific transgene expression with affinity purification of translating ribosomes. TRAP can be used to study the cell type–specific mRNA profiles of any genetically defined cell type, and it has been used in organisms ranging from Drosophila melanogaster to mice and human cultured cells. Unlike other methodologies that rely on microdissection, cell panning or cell sorting, the TRAP methodology bypasses the need for tissue fixation or single-cell suspensions (and the potential artifacts that these treatments introduce) and reports on mRNAs in the entire cell body. This protocol provides a step-by-step guide to implement the TRAP methodology, which takes 2 d to complete once all materials are in hand.
MIT Department
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Picower Institute for Learning and Memory
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Creative Commons Attribution-Noncommercial-Share Alike
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DOI of Published Version
https://doi.org/10.1038/nprot.2014.085
