Global mapping of translation initiation sites in mammalian cells at single-nucleotide resolution

Understanding translational control in gene expression relies on precise and comprehensive determination of translation initiation sites (TIS) across the entire transcriptome. The recently developed ribosome profiling technique enables global translation analysis, providing a wealth of information about both the position and density of ribosomes on mRNAs. Here we present an approach (global translation initiation sequencing, GTI-seq), by applying in parallel ribosome E site translation inhibitors lactimidomycin (LTM) and cycloheximide (CHX), to achieve simultaneous detection of both initiation and elongation events on a genome-wide scale.