Fig. 3.

Detection of dual-sgRNAs/Cas9-mediated GHR modification in piglets. a, b Picture of GHRKO pigs showing their small stature and normal fertility. c Picture of the live GHRKO fetus that was used to establish cell lines for recloning. d PCR products harboring the targeted region of GHR amplified from selected piglets. e Detection of dual-RNAs/Cas9-mediated on-target GHR cleavage by the T7ENI cleavage assay in piglets. P1–P3 piglets were cloned from the C3 single-cell colony; P4–P7 piglets were cloned from the C6 single-cell colony. (M, DNA maker DL2000; C, colony; WT, wild-type PCR product digested by T7ENI). f Modified GHR sequences detected in piglets and fetuses. F1 and F2 indicate the two fetuses cloned from the C6 cell colony. P4–P15 were cloned piglets from C6 cell colony