DESIGN OF mRNA SEQUENCES TO CONTROL CO-TRANSLATIONAL FOLDING OF PROTEINS

摘要

This invention relates to optimized heterologous production of properly folded and functional proteins. The present invention provides systems and methods involving determination of the optimal mRNA sequence, based on the underlying rates at which codons are translated and folding kinetic of nascent-protein, that maximizes co-translational protein folding of domains in order to maximize the proper folding and quality of the protein produced. The codon translation rates can be determined in a number of ways, including theoretical estimation and, preferably, through experimental data, such as ribosome profiling. The determination of an optimal mRNA sequence through the utilization of codon translation rates under a particular set of conditions allows for application of the method irrespective of the organism from which the protein in question was originally derived.

主权项

1. A method of enhancing heterologous protein production in a cell comprising:
(a) providing a protein having a known amino acid sequence; (b) providing conditions for expression of said protein; (c) providing the rates of translation for said conditions of expression; (d) providing the conformational states said protein can populate; (e) providing the rates of interconversion among the states in (d); (f) selecting a desired conformational state from (d) for production of said protein; (g) producing an optimized mRNA sequence for said protein in the selected desired conformational state using an algorithm that accounts for (a)-(e).