Higher Efficiency of Transgenic Mouse Production Achievedby Double-pronuclear Microinjection

Objective To establish an efficient technical platform for genetically-engineering animal model researches. Methods Using Kunming mice as the laboratory animals, DNA molecules (K14/hCTLA4-Ig expression vector) at a concentration of 10 ng/μl was transferred into male and female pronuclei of mouse eggs by double-pronuclear microinjection, while single-pronuclear microinjection was performed as control. Cleaved injected eggs were transferred into oviducts of synchronized recipient mice. Offsprings were screened by PCR. Results In the double-pronuclear microinjection group: the cleavage rate of injected eggs was 45.0% (108/240), significantly lower than that of single-pronuclear injection group (52.3%); the survival rate in vivo of transferred eggs was 18.5% (20/108), with no significant difference compared to that of control group (16.7%); the transgenic rate of founder mice and total injected eggs was 27.8% and 2.08%, respectively, which was significantly higher than that of control (12.5% and 1.2%). Conclusion By double-pronuclear injection, transgenic mouse production efficiency can be effectively improved.