Fig. 1: Knock-in efficiency of dsDNA donor positively correlates with MMEJ repair patterns in mouse embryos.

A Schematic of three sgRNAs targeting the mouse Actb and Cdx2 loci. “Stop” marks the stop codon. “d” indicates the distance (in bp) from the sgRNA cut site (blue triangle) to the donor DNA insertion site (green triangle), as shown. B Workflow of CRISPR reagent injection and embryo repair pattern analysis. Zygote, blastocyst, and CRISPR icons were created in BioRender. Hongyu, C. (2025) https://BioRender.com/qxdvotz. C Indel frequencies for different sgRNAs at Actb and Cdx2 loci. Each dot represents a blastocyst; n values and exact p-value are shown. Data represent mean ± SEM. Two-sided Kruskal-Wallis test was used. D Pie charts show DNA repair pattern distributions and NHEJ/MMEJ ratios per sgRNA. E Schematic of the dsDNA knock-in strategy. Figure was partially created in BioRender. Hongyu, C. (2025) https://BioRender.com/qxdvotz. F Representative fluorescence images and quantification of mCherry knock-in at Actb and Cdx2 loci (scale bars, 100 μm). Each dot: n = 3 experiments with > 20 embryos per condition. Bars: mean ± SEM. Two-sided unpaired t-tests; p-value in figure. G Correlation between knock-in efficiency and indel/MMEJ/NHEJ repair ratios. Each dot: one sgRNA. Red line: linear regression. R² and p-value shown in figure. Full statistics in Source Data. Source data are provided as a Source Data file.