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bioRxiv · 10.1101/2020.06.25.169177

Custom-Designed, Degradation-Resistant Messenger RNAs in Yeast

Franklin, A. L.·Macfadden, A.·Kieft, J. G.·Hesselberth, J.·Chapman, E. G.
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Abstract

Virally-derived RNA structures provide useful tools for engineering messenger RNA (mRNA) transcripts with unique properties. Here we show that 5[->]3 exoribonuclease-resistant RNA structures (xrRNAs) from Flaviviruses can be used to protect heterologously-expressed messenger RNAs (mRNAs) from 5[->]3 degradation in Saccharomyces cerevisiae (budding yeast). Installation of xrRNAs ahead of a downstream internal ribosome entry site (IRES) leads to the accumulation of partially-degraded mRNAs designed to undergo cap-independent translation. In a yeast strain favoring cap-independent translation, this strategy led to a 30-fold increase in the enzymatic activity of lysates obtained from cells expressing degradation-resistant reporters. Overall, these finding demonstrate the possibility of coupling multiple viral RNA structures together to enhance protein expression.

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Franklin, A. L., Macfadden, A., Kieft, J. G., Hesselberth, J., Chapman, E. G.. 2020-06-26. Custom-Designed, Degradation-Resistant Messenger RNAs in Yeast. https://doi.org/10.1101/2020.06.25.169177

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