programmed ribosomal frameshift Raw sequence reads

Protein translation in eukaryotes is reprogrammed through diverse molecular mechanisms to expand the coding capacity of individual genes. One such mechanism, termed -1 programmed ribosomal frameshifting (-1 PRF), utilizes cis-acting signals within mRNA to redirect ribosomes into a new translation reading frame, thereby giving rise to an alternative protein product. We establish -1 PRF as a powerful framework for engineering small molecule-responsive RNA switches that control protein synthesis in eukaryotes. Efficient -1 PRF stimulatory elements were discovered by in vitro selection and characterized by high-throughput sequencing; then, ligand-responsive -1 PRF switches were constructed by coupling -1 PRF stimulatory elements to RNA aptamers using rational design and in vivo directed evolution.