Directed evolution of orthogonal transcription engine for programmable gene expression in eukaryotes

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.

本数据集收录了在融合酶控制下，由T7 RNA聚合酶和capping酶NP868R变体构成的酵母菌酿酒酵母（Saccharomyces cerevisae）和HEK293T细胞中荧光表达诱导的折叠变化。图1c展示了经过多轮选择后，与野生型相比的NPT7进化变体的特性。在半乳糖诱导后，报告基因表达的折叠诱导。图1f展示了使用梯度半乳糖诱导对野生型NPT7、v433和v443的报告基因表达的特性。图h展示了在基因组中整合以及作为多拷贝酵母质粒编码的情况下，由Gal启动子控制的报告基因表达。图2b展示了v433和v443菌株的所有报告基因表达的特性。图2d展示了突变T7 RNAP启动子驱动的报告基因表达呈现出与预测启动子强度一致的分级反应。图2f-）展示了v433和v443菌株中每个荧光载体的相对表达量相对于BFP。图3b展示了六个v443变体对六个报告构建体的活性特性。图4b展示了在转染每个报告质粒后，野生型NPT7、v433和v443在HEK293T细胞中的活性特性。