Synonymous codon usage regulates translation initiation

Ribo-seq with firefly luciferase optimality reporters to determine whether incomplete translation and/or inhibited translation initiation is responsible for the reduced ribosome occupancy on nonoptimal transcripts. Performed Ribo-seq, with paired RNA-seq, on HEK293T cells transfected with either optimal or nonoptimal reporters. Experiments were performed in triplicate.