Journal: bioRxiv

Article Title: Dual function of ERH in primary miRNA biogenesis

doi: 10.1101/2025.09.23.678008

Figure Lengend Snippet: A . Scaled scheme of the human DGCR8 mRNA, with individual exons depicted as numbered boxes. The ORF is marked in dark grey. B . Expression of pre-miR-3618 and miR-1306 as quantified by RT-PCR. Individual datapoints are illustrated as colored circles and triangles depending on the sgRNA (n=3+3). C . PCR-quantified expression of depicted miRNAs and mRNAs in control and miR-3618 KO cells. D , E . Expression of DGCR8 and DROSHA mRNAs in SAFB1/2 and ERH KO cells transduced with different sgRNAs, and the corresponding western blot analysis for protein expression and a quantification thereof. F . Expression of miR-1306 and DGCR8 RNA as well as protein levels in SAFB1/2 KO cells reconstituted with the depicted SAFB2 variants. Numbers indicate p-values.

Article Snippet: BoxB constructs for the mutated variants of miR-15a and -181b were generated based on the RNA motif plasmid (Addgene #107253, kindly provided by Paul Khavari) of the RaPID system , and were expressed with the BoxB-miRNA cassette placed in the 3’-UTR of a cDNA encoding a intracellular tail-deletion mutant of the cell surface marker CD8.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Transduction, Western Blot

Journal: bioRxiv

Article Title: Dual function of ERH in primary miRNA biogenesis

doi: 10.1101/2025.09.23.678008

Figure Lengend Snippet: A . Model for the temporal sequence of cluster assistance, beginning with Microprocessor recruitment to the helper, followed by helper processing and subsequent transfer of DROSHA/DGCR8 to the recipient. B . Predicted structure of the cluster assistance-recipient pri-miR-181b and its seed mutant used for the tethering experiments. C . Control experiments for the Microprocessor tethering as in , indicating no miRNA reporter repression in the absence on any BoxB aptamer sites in the substrate RNA. D . Corresponding experiment to , using tethering of λN-DGCR8 to miR-15a mut and its respective reporter as a readout. Histogram overlays and the bar graph depict changes in dsRed fluorescence in consequence of SAFB1/2 and ERH knockout. Data points are shown as circles and triangles for the individual sgRNAs (n=3+3). Numbers indicate p-values.

Article Snippet: BoxB constructs for the mutated variants of miR-15a and -181b were generated based on the RNA motif plasmid (Addgene #107253, kindly provided by Paul Khavari) of the RaPID system , and were expressed with the BoxB-miRNA cassette placed in the 3’-UTR of a cDNA encoding a intracellular tail-deletion mutant of the cell surface marker CD8.

Techniques: Sequencing, Mutagenesis, Control, Fluorescence, Knock-Out