tsRibo-Seq: An Approach to Zoom In into Translation of Single Transcripts

Ribo-Seq (ribosome profiling) enables global assessment of translation efficiency, yields and fidelity. However, low-abundance transcripts receive little coverage and fall outside of the Ribo-Seq detection limit. We developed a new twist of the Ribo-Seq approach, termed transcript specific Ribo-Seq (tsRibo-Seq) to specifically detect lowly abundant messenger RNAs (mRNAs). This approach yields unprecedented depth in the coverage of individual low-abundance transcripts allowing identification of slow and fast translating regions with nucleotide resolution. Overall design: Ribosome Profiling of CFBE cells 48h after transfection with wild-type CFTR