Directed evolution of orthogonal transcription engine for programmable gene expression in eukaryotes
Description
This data set contains fold induction for fluorescence expression in saccharomyces cerevisae and HEK293T cells under the control of fusion enzymes composed of T7 RNA polymerase and capping enzyme NP868R variants. Figure 1c - Characterization of evolved variants of NPT7 relative to wild-type after rounds of selection. Fold induction of reporter expression following galactose induction. Figure 1f - Characterization of reporter expression using graded galactose induction for WT NPT7, v433 and v443. For Figure h) Reporter expression under the control of the Gal promoter assayed both integrated in the genome as well as encoded as a multi-copy yeast plasmid. For Figure 2b - Characterization of all reporter expression for both v433 and v443 strains. Figure 2d - Reporter expression from mutant T7 RNAP promoters shows a graded response consistent with the predicted strength of the promoters. Reporter expression from mutant T7 RNAP promoters shows a graded response consistent with the predicted strength of the promoters. Figure 2f- ) Relative expression of each fluorescent cargo relative to BFP for both v433 and v443 strains. Figure 3b - Characterization of all six v443 variants activity against the panel of six reporter constructs. Figure 4b- Activity of WT NPT7, v433 and v443 were characterized in HEK293T cells after transfection with each reporter plasmid.
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Steps to reproduce
Detailed methods have been provided in Methods section of the manuscript titled - Directed evolution of orthogonal transcription engine for programmable gene expression in eukaryotes currently in iScience.