Fig. 3: Identification of Gua-responsive HDV ribozymes by amplicon-seq. | Nature Communications

Fig. 3: Identification of Gua-responsive HDV ribozymes by amplicon-seq.

From: High-throughput identification of synthetic riboswitches by barcode-free amplicon-sequencing in human cells

Fig. 3

a Secondary structure of the Gua-HDV ribozyme library design. Blue nucleotides indicate the randomized motif. The arrowhead indicates the cleavage site. C/U mutation at the indicated position renders the ribozyme inactive. b Guanine riboswitch construct frequency analysis of the original library. c Volcano plots displaying the significance (FDR) of changes in riboswitch abundance as a function of the change in expression (log2 fold change). d Selected hits from the Gua-HDV riboswitch library screen, randomized sequence, mean counts per million (CPM), fold change (FC) and false-discovery rate (FDR) at 30, 100, and 300 µM guanine, respectively. The bar plots show the raw counts under untreated, 30, 100, and 300 µM µM Gua-stimulated conditions (four groups from left to right, n = 8 replicates each). e Functional hit validation in HEK-293 cells at increasing guanine doses [µM] (n = 3 replicates, mean ± s.d.); HDV-act/inact: constitutively active/inactive ribozyme control. f Maximal fold changes and p-values measured upon stimulation vs. 0 µM guanine for all tested constructs. *p < 0.05, **p < 0.01, ***p < 0.001 (unpaired T-Test, two-tailed). Source data are provided as a Source Data file.

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