Fig. 4

Validation of indel in MBCD-SMGE derived blastocyst by sequence trace decomposition method using inference of CRISPR edits (ICE) software. A Overview of the tracking process of targeted indel mutations in MBCD-SMGE derived blastocysts. B Gel-electrogram image shows isolated GOI from GFP-positive blastocyst obtained from sperm incubated in c-TYH with 2 mM MBCD and 20 ng/µl pgRNA-Cas9 (L2–L5) and control blastocyst (L6). L1: No-template control, and L: 100 bp DNA ladder. C-F: Output of computational analysis by ICE. C The relative contribution of the wild-type and edited sequence in four GFP-positive blastocyst samples (1-3: non-edited samples, 4: edited sample). D Sanger sequencing view displays the edited (a), non-edited (b, c, and d), and wild-type (control) sequences in the vicinity of the guide region. The guide sequence is represented by the black region that is underlined horizontally. E The indel plot of the edited blastocyst reveals the estimated distribution of indels across the entire genome of the blastocyst. F The discordance plot of the edited blastocyst provides details about the degree of similarity per base between the wild-type (control) and the edited sample within the specific region surrounding the cut site (inference window)