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An inducible CRISPR interference library for genetic interrogation of Saccharomyces cerevisiae biology

机译:一种诱导诱导酵母菌遗传思想生物学遗传询问的诱导乐谱文库

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摘要

a Schematic of amPL43 expression vector for inducible CRISPRi library in S. cerevisiae. b The expression fold change of each target: ERG25 (three replicates), ERG11 (five replicates), and Sec14 (four replicates), as a result of presence of sgRNA, without induction after 24 hours, as measured by qPCR. The mean for each sample is represented by a solid line. c The expression fold change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. d Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. e Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. f Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.
机译:酿酒酵母诱导型CRECRI文库的催化卷积载体的示意图。 b作为SGRNA的存在的结果,每个目标的表达倍换:ERG25(三次重复),ERG11(五重复),ERG11(五重复)和SEC14(四重复),但通过QPCR测量,在24小时后没有诱导。每个样品的平均值由实线表示。 C通过QPCR计算每个目标的表达折叠变化:ERG25(三次重复),ERG11(三重复),ERG11(三次重复)和SEC14(两次重复),随着QPCR的时间计算。每个样品的平均值由实线表示。 D示意图描绘了PTA1和ERV46的基因组区域。靶向两种基因之间的区域的GRNA,取决于它们对每个基因的邻近,可能影响两个基因。 E直方图描绘了两个图书馆重复的每个基因的GRNA数量。虚线的蓝线表示中位数:〜6。 F散射曲线描绘CRISPRI文库选择生物重复之间的每GRNA读数的频率。每个对报告Pearson相关性R值。

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