Figure 3. Development of reconstructed transgenic embryos.

shRNA recombinant lentivirus vector RNAi-VP4 was transfected into bovine fetal fibroblast cells using Lipofectamine 2000, transgenic bovine fetal fibroblast cells were screened by FACS, all of them expressed eGFP (A), and confirmed by PCR with primer pair LT1/LT2 (B) and DNA sequencing. shRNA transgenic cells were transferred into enucleated oocyte cytoplasts, and transgenic blastocysts were screened based on their development and eGFP expression (C).