Figure 1.

Optimization of the jetCRISPR concentration for gene editing of MSTN. (A) Blastocyst formation rates of ZP-intact and ZP-free embryos without RNP transfection. (B) Frequency of gene editing in the target regions of blastocysts derived from the embryos treated with jetCRISPR, Cas9 protein, and gRNA. Gene editing of blastocysts was determined by Sanger sequencing and TIDE. The percentage of blastocysts with gene editing was defined as the ratio of the number of gene edited blastocysts to the total number of blastocysts examined. (C) Mutation efficiency in gene-edited blastocysts. Editing efficiency was defined as the proportion of indel mutation events in blastocysts carrying mutations. Heterozygous without WT: blastocysts carrying multiple types of editing but no WT sequences, Heterogeneous with WT: blastocysts carrying mosaic mutation or heterozygous mutation carrying more than one type of mutation and the WT sequence, and monoallelic mutation. (D) Blastocyst formation rates of embryos treated with various concentrations of jetCRISPR. Each bar represents the mean ± SEM. Four replicate trials were carried out and the numbers in parentheses indicate the total number of oocytes (A,D) and examined blastocysts (B,C). Percentages of blastocysts carrying mutations in target genes were analyzed using chi-squared tests (B). *p < 0.05.