Underscoring the value of measuring gene expression at the level of translation has led to the development of ribosome-profiling techniques. Ribosome profiling is a direct method for determining the precise mRNA region being read by the ribosome, to examine the transcripts associated with the translation machinery directly. Ribosome profiling providing a cellular snapshot of protein production, with the potential to inform nearly every domain of biological and clinical research. Compared with traditional RNA-Seq experiments, ribosome profiling is designed specifically to measure mRNA involved inactive translation.
CD genomics provides a ribosome-profiling strategy that is based on the deep sequencing of ribosome-protected mRNA fragments and enables genome-wide investigation of translation with sub cordon resolution. Using nuclease digestion, the ribosome position and the translated message can be precisely determined by analyzing the protected ~30-nt area of the mRNA template.
Fig.1 Ribosome-protected mRNA
Application of Ribosome Profiling
Ribosome Profiling Workflow
Using nuclease digestion to isolate nuclease-resistant ribosome, the translated message can be precisely determined by analyzing the protected 30 nucleotides during translation.