Fig. 3
From: CRISPR-Cas9 genome editing induces megabase-scale chromosomal truncations
Unique DSB impairs chromosomal integrity. a, b DNA-FISH assay using (a) UROS-framing probes (respectively −4.6 Mb upstream and + 4.4 Mb downstream from UROS locus) or (b) Chr10-specific sub-telomeric probes for NT HEK293T cells or cells transfected with nuclease only, nuclease + ssODN or nickase + ssODN. For the two couples of probes, green (G) and orange (O) fluorescent probes are respectively upstream and downstream of the UROS gene. In this way, Chr10q terminal truncation is denoted by loss of orange signals. (Left) quantification of cells with 3 O/3 G, 2 O/3 G or 1 O/3 G signals. (Right) Illustrative DNA-FISH results for HEK293T. Statistical significance is inferred using two-sided chi-square test (versus NT cells). ns, not significant; *p < 0.05 **p < 0.01. c Array-CGH on truncated clones. First, HEK293T were transfected with nuclease and analyzed by FISH using UROS-framing probes. Transfected cells were subcloned and 3 out of 10 clones were identified as 2 O/3 G. Chromosome 10 integrity of clones #8 and #10 was evaluated by array-CGH. Deletion in #10: arr[GRCh37] 10q26.2q26.3(127516127_135404523)x2. Duplication and deletion in clone #8. arr[GRCh37] 10q24.1q26.2(95667790_127496056)x3~4,10q26.2q26.3(127516127_135404523)x2. d DNA-FISH assay using UROS-framing probes for primary wild-type fresh hFF (human Foreskin Fibroblasts) (hFF), immortalized with hTERT (hFF hTERT) or TP53−/− immortalized fibroblasts (hFF hTERT TP53−/−), NT or transfected with nuclease. Quantification of 2 O/2 G and 1 O/2 G signals percentages. For (a, b, c), source data are provided as a Source data file
