Reprogramming eukaryotic translation with ligand-responsive synthetic RNA switches
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Protein synthesis in eukaryotes is regulated by diverse reprogramming mechanisms that expand the coding capacity of individual genes. One such mechanism is programmed ribosomal frameshifting (PRF). In this work, efficient PRF stimulatory RNA elements were discovered by in vitro selection, and then ligand-responsive switches were constructed by coupling PRF stimulatory elements to RNA aptamers using rational design and directed evolution. Motif discovery was enabled by the methodological novelty of deep sequencing an initially randomized library of RNA sharing a certain pseudoknot scaffold that had undergone multiple rounds of in vitro selection for PRF. This approach led to a rich characterization of precise pseudoknot geometries that can facilitate translation reprogramming, an area with great potential for synthetic biology.
“Reprogramming eukaryotic translation with ligand-responsive synthetic RNA switches.”
AUTHORS: Anzalone AV, Lin AJ, Zairis S, Rabadan R, Cornish VW.
LINK TO PUBLICATION:
Nat Methods. 2016 Mar 21. doi: 10.1038/nmeth.3807.