Developmental competence and mutation efficiency on
microinjection mediated gene editing
To determine optimal conditions, including Cas9 mRNA, and sgRNA
concentrations for producing mutations in bovine blastocysts, four
experimental conditions were used, then developmental competence and
mutation efficiency were investigated in the blastocyst stage following
each condition (Fig 1C, D, and E). There was no significant difference
in cleavage rate across all 4 groups (data not shown). However, in the
RNA4X group a diminished blastocyst formation rate was observed in
comparison to the control group (Fig 1D). In terms of mutation rate, the
RNA2X group (81.3 ± 17.2%) had the highest mutation compared the other
3 groups (WT: 0%, RNA1X: 33.3 ± 16.0%, RNA4X: 50.0 ± 28.5%) (Fig 1E).
Thus, the RNA2X condition was selected as the optimal condition for the
further production of MSTN mutated cattle.