Journal: Nature Communications
Article Title: Unveiling the cut-and-repair cycle of designer nucleases in human stem and T cells via CLEAR-time dPCR
doi: 10.1038/s41467-025-65182-4
Figure Lengend Snippet: Characterisation of aberrations induced by repair inhibitors on Cas9 edited HSPCs targeting the CCR5 locus. a Summary comparison between CLEAR-time dPCR (top) and ICE (bottom) 1, 3, and 14-days post-editing. Rel. indels = relative indels. For CLEAR-time dPCR, data are shown as mean ± s.d. For ICE, data are shown as n = 1. b Quantification of sequence trimming around the cleavage site in CCR5 edited HSPCs 1, 3, and 14-days post-editing. Data are shown as mean ± s.d. c CLEAR-time dPCR summary of aberrations induced by repair inhibitors on Cas9 edited iPSCs (top) and HSPCs (bottom) targeting the CD34 locus with and without an ssODN 3, 7, and 14-days post-editing. Data are shown as mean ± s.d. d Quantification of sequence trimming around the cleavage site of CD34 edited iPSCs (top) and HSPCs (bottom) 1, 3, and 14-days post-editing. Data are shown as mean ± s.d. e Comparison between ICE (I) and CLEAR-time dPCR (Ct) in Cas9 edited T cells (top), HSPCs (middle), and iPSCs (bottom) targeting the CCR5 locus with and without AAV transduction and AZD7648 treatment 4-days post-editing. Data are shown as mean ± s.d. ( n = 3 technical replicates of 2 independent donors per cell type). All data represents n = 3 technical replicates unless stated otherwise. b , d Two-way ANOVA with Tukey post-hoc test., n.s.= non-significant, **** p < 0.001. Source data are provided as a Source Data file.
Article Snippet: DNA-PK inhibitor AZD7648 and POLQ inhibitor ART558 were purchased from MedChemExpress or Cambridge Bioscience.
Techniques: Comparison, Sequencing, Transduction
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