TRAP-Seq
TRAP-Seq
Targeted purification of polysomal mRNA (TRAP-Seq) maps translating mRNAs under various conditions. In this method, tagged ribosomal proteins are expressed in cells. The tagged ribosomal proteins are purified and the RNA isolated. The RNA is reverse-transcribed to cDNA. Deep sequencing of the cDNA provides single-base resolution of translating RNAs.
Pros:
- Allows detection of translating RNAs
- RNAs translated by specific targeted ribosomes can be assessed
- No prior knowledge of the RNA is required
- Provides a genome-wide RNA screen
Cons:
- Not as specific as more recently developed methods, such as Ribo-Seq
- Nussbacher J. K., Batra R., Lagier-Tourenne C. and Yeo G. W. RNA-binding proteins in neurodegeneration: Seq and you shall receive. Trends Neurosci. 2015;38:226-236
- Maze I., Shen L., Zhang B., et al. Analytical tools and current challenges in the modern era of neuroepigenomics. Nat Neurosci. 2014;17:1476-1490