Journal: Genome Biology

Article Title: Recruitment and rejoining of remote double-strand DNA breaks for enhanced and precise chromosome editing

doi: 10.1186/s13059-025-03523-8

Figure Lengend Snippet: Living-cell images present homologous donor template bringing broken chromosomes ends into close proximity. a Diagram of donor induced translocation between Chr3 and Chr13 chromosomes. In U2OS cells, Chr3Rep and Chr3Rep were labeled with CRISPRainbow system. Donor plasmid was labeled by ParB-ParS system. SpCas9/sgRNA induces DSB between two marker sites by nuclear transfection (16 h after delivery of the DNA site marker system). b Fluorescence images of separation and reconnection of chromosome C3 and C13 sites. Red fluorescence represents the C3 site, and purple fluorescence represents the C13 site. Donor plasmid is shown by GFP fluorescence. HRMR, with homologous donor. CK, with non-homologous donor. The yellow boxes represent the DNA sites indicated by the arrows. Scale, 5 µm. c The distance between C3 and C13 sites was measured before and after chromosome reconnection in different cells

Article Snippet: At 72 h after transfection, genomic DNA was extracted using a DNA Extraction Kit (Tiangen).

Techniques: Translocation Assay, Labeling, Plasmid Preparation, Marker, Transfection, Fluorescence