Journal: Communications Biology

Article Title: Plasmid2MC: efficient cell-free generation of high-purity minicircle DNA for genome editing in mammalian cells

doi: 10.1038/s42003-025-09157-7

Figure Lengend Snippet: a Diagram of the source plasmid and the resulting mcDNA with excluded bacterial backbone, used in this validation edit. b Plasmidsaurus sequencing histogram showing the high quality of the mcDNA product. c A circular vector for the expression of gRNA for base editing targeting, using the method developed earlier in our laboratory . d Base editing (dCas9 ABE8e) A → G conversion rates in HEK293T cells for the first three sites from . e dCas9 ABE8e editing A → G conversion rates in two mESC sites. Colors: mcDNA (green) and the original plasmid (orange). Guide sequences for the sites are followed by PAM (in bold red font). In plots ( d , e ), standard deviation (SD) values are indicated with the red line range. Where not visible, the SDs fall within the spread of the data points.

Article Snippet: Fig. 4 Validation of mcDNA performance in ABE8e dCas9 base editing. a Diagram of the source plasmid and the resulting mcDNA with excluded bacterial backbone, used in this validation edit. b Plasmidsaurus sequencing histogram showing the high quality of the mcDNA product. c A circular vector for the expression of gRNA for base editing targeting, using the method developed earlier in our laboratory . d Base editing (dCas9 ABE8e) A → G conversion rates in HEK293T cells for the first three sites from . e dCas9 ABE8e editing A → G conversion rates in two mESC sites.

Techniques: Plasmid Preparation, Biomarker Discovery, Sequencing, Expressing, Standard Deviation

Journal: Communications Biology

Article Title: Plasmid2MC: efficient cell-free generation of high-purity minicircle DNA for genome editing in mammalian cells

doi: 10.1038/s42003-025-09157-7

Figure Lengend Snippet: a Schematic showing the design of HITI constructs. Blue pentagon, Cas9/gRNA target sequence in the host genome and HITI mcDNA. HygR: Hygromycin-resistant gene. Black square: non-functional random DNA sequence. b Schematic of HITI experiment with antibiotic selection to enrich cells with the correct HITI-mediated integration before subsequent analysis. c Flow cytometry analysis of cells treated with HITI or control (untransfected cells). Percentage of positive population is shown within the gates. d Gel electrophoresis of PCR overlapping the mKate gene and either the ACTB or GAPDH genes. HITI - shows bands for edited samples, while no bands are visible for unedited Controls. See Supplementary Fig. for additional analysis and unprocessed gel electrophoresis photos.

Article Snippet: Fig. 4 Validation of mcDNA performance in ABE8e dCas9 base editing. a Diagram of the source plasmid and the resulting mcDNA with excluded bacterial backbone, used in this validation edit. b Plasmidsaurus sequencing histogram showing the high quality of the mcDNA product. c A circular vector for the expression of gRNA for base editing targeting, using the method developed earlier in our laboratory . d Base editing (dCas9 ABE8e) A → G conversion rates in HEK293T cells for the first three sites from . e dCas9 ABE8e editing A → G conversion rates in two mESC sites.

Techniques: Construct, Sequencing, Functional Assay, Selection, Flow Cytometry, Control, Nucleic Acid Electrophoresis