We used ICR and B6D2F1 (C57BL/6 x DBA2 F1) female mice in this study. The ICR strain was mainly used for determining the optimal conditions for electroporation, and the B6D2F1 strain was used for genome editing.
Fertilized eggs were collected from the oviducts of E0.5 (12 hours after the midpoint of the day of vaginal plug) ICR or B6D2F1 females naturally intercrossed with males of the same strain. The covering cumulus cells were removed by incubating in 1% hyaluronidase/M2 medium (Sigma). For the genome editing experiments targeting H2b-mCherry, the eggs were obtained from B6D2F1 females intercrossed with R26-H2b-mCherry males. The collected eggs were pre-cultured in mWM medium until electroporation.
Fertilized eggs were collected from the oviducts of E0.5 (12 hours after the midpoint of the day of vaginal plug) ICR or B6D2F1 females naturally intercrossed with males of the same strain. The covering cumulus cells were removed by incubating in 1% hyaluronidase/M2 medium (Sigma). For the genome editing experiments targeting H2b-mCherry, the eggs were obtained from B6D2F1 females intercrossed with R26-H2b-mCherry males. The collected eggs were pre-cultured in mWM medium until electroporation.
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