Fig. 1: scSNP-DNAseq design to assay CRISPR-induced ON-target genotoxicity in chromosomes 10 and 11 (Chr10/11), and method validation for LOH quantification. | Nature Communications

Fig. 1: scSNP-DNAseq design to assay CRISPR-induced ON-target genotoxicity in chromosomes 10 and 11 (Chr10/11), and method validation for LOH quantification.

From: Single-cell multiplex approaches deeply map ON-target CRISPR-genotoxicity and reveal its mitigation by palbociclib and long-term engraftment

Fig. 1: scSNP-DNAseq design to assay CRISPR-induced ON-target genotoxicity in chromosomes 10 and 11 (Chr10/11), and method validation for LOH quantification.The alt text for this image may have been generated using AI.

a Design of home-made scSNP-DNAseq library in Chr10/11 focusing on frequent SNPs. b Mapping of amplicon densities around ABRAXAS2, UROS and HBG1/2p breakpoints. c Method validation using hFAMRed system. Heterozygous UROS +/− hFFs switched from non-fluorescent to fluorescent in case of CRISPR-induced Chr10q telomeric LOH encompassing UROS. Chr10 is represented with ABRAXAS2 and the five SNPs used to detect LOHs by scDNA-seq. ABRAXAS2 targeting by CRISPR-Cas9 induced fluorescent cells 15 days after editing (0.1–0.2%). Fluorescent cells were sorted and cultured for 2 months to obtain enough cells before analysis by scSNP-DNAseq and Mosaic 3 software. d Histogram of SNP profiles in unedited/edited fluorescent hFFs (fluo+). e different SNP profiles identified by sc-SNP-DNA seq (WT, LOH 10qI, LOH 10qII). Created in BioRender. Bedel, A. (2025) https://BioRender.com/3fwjwd9.

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