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Fig. 4 | Genome Biology

Fig. 4

From: Effective control of large deletions after double-strand breaks by homology-directed repair and dsODN insertion

Fig. 4

Insertion of double-stranded oligonucleotides reduces large deletions in T cells. a Schematic of dsODN insertion into DSBs with Cas9-gRNA RNPs and dsODN. dsODN is inserted in both forward and reverse orientations. b Frequencies of small indels and 34-bp dsODN insertions at three loci in T cells. The numbers in the bars indicate the mean small indels or dsODN insertion efficiencies. Small indels (NHEJ) and dsODN insertions (treated as HDR by CRISPResso2) were determined by amplicon sequencing and CRISPResso2 analysis. c, e Insertions with 28, 29, or 34 bp-dsODN decrease deletion indexes in T cells. Deletion indexes were assessed by long PCR, nanopore sequencing, Minimap2 alignment, and Samtools analysis. d Frequencies of small indels and dsODN insertions after editing with RNP and 28- or 29-bp dsODN. Error bars represent the mean ± SEM of n = 3–6 independent experiments. The data in b–e were statistically analyzed by a two-way ANOVA test. Adjusted p values were indicated. “ns” means no significance (p > 0.05)

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