Fig. 4.

Efficiently generating three gene mutations in Bama miniature pig genome by somatic cell nuclear transfer (SCNT). a Procedure for pig SCNT using Fetal Bama miniature pig fibroblasts. Spindle–chromosome complex (arrowhead) in a porcine MII oocyte before (I) and after (II) removal; an oocyte with a slit in the zona pellucida (III, arrowhead) induced by a beveled glass pipette, and the Fetal Bama miniature pig fibroblasts (arrowhead) before (IV) and after (V) injection into the perivitelline space; the blastocysts developed from injected embryos (VI), which were produced by SCNT using fetal fibroblasts. b Photo of F₀ piglet at 4 weeks after birth. c Representative sequencing chromatograms at the GGTA1, B4galNT2, and CMAH targets of WT and #502. The mutation amino acids are shown in the red boxes. Detection of aGal (d), Sd(a) (e), and Neu5Gc (f) expression profiles on the surface of fibroblasts from WT and base mutation pig (#502) by IFA, respectively. The nucleus in the PFFs is shown in blue dots, and the aGal, Sd(a), and Neu5Gc on the surface of PFFs are shown in red. Bars represent mean ± SD, #502: n = 3, WT: n = 3. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001