Quantifying the impact of codon usage on mRNA translation efficiency through massive parallel reporter assays.

We introduced synonymous mutations randomly into two regions of EGFP. After culturing in HEK293 cells, we quantified the translation efficiency for each sequence by combining sucrose density gradient centrifugation and cell flow cytometry, followed by high-throughput sequencing. This allowed us to explore the impact of codon usage combinations on the translation process.