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.
KeywordsmRNA Cell type Posttranscriptional regulation Transcriptome
We thank J. Bailey-Serres for sharing protocols. This work was supported by the National Natural Science Foundation of China Grants 31222033 and 31171159, by the National Basic Research Program of China (973 Program) Grant 2012CB910902, and by the Hundred Talents Program of CAS.