动物造模 z-bio 2021-07-02 273

　　Objective: To explore the feasibility of injecting Cas9 after in vitro fertilization in freeze-thaw mice.

　　Method: After IVF with fresh C57BL/6J mouse eggs, prokaryotic embryos (single-cell embryos) and 2-cell embryos are frozen by cryotube method (EFS20/40), resurrected the next day, and cultured later. I will. Compare the recovery rate and survival rate of frozen-thawed prokaryotic embryos and 2-cell embryos. Cas9 and the diluted cytoplasm were injected into the embryos of some freeze-thaw prokaryotes and fresh prokaryotes at the same time, cultured into 2 cells, and compared the survival rate after injection and the incidence of 2 cells.

　　Result: Is the recovery rate of B6 mouse prokaryotic embryos after freezing and thawing 92.5? Survival rate 92.8? 2 cell embryo recovery rate 90.5? Survival rate 95.8? A set of data, there is a significant difference between the two (Pu003c0.05). Is the survival rate of fresh prokaryotic embryos after Cas9 injection 92.7? Is the survival rate of the blank group 97.5? Is the survival rate of pre-frozen and thawed embryos 82.6 after Cas9 injection? Is the survival rate of the blank group 97.5? 92? The difference between the freeze-thaw group and each group is different. Important (P0.05). There was no significant difference in the incidence of 2-cell embryos after injection (P u003c0.05).

　　Conclusion: Freeze-thawed prokaryotic embryos can be used for Cas9 microinjection.