Title: GENE INTEGRATION EFFICIENCY AND ZYGOTES SURVIVING IN MICE AT PRONUCLEAR MICROINJECTION
Abstract:By applying the microinjection into pronuclear of murine zygotes we demonstrated the successful integration of gene LOC 78634 construction at a comparative level efficiency and transgenesis. The study...By applying the microinjection into pronuclear of murine zygotes we demonstrated the successful integration of gene LOC 78634 construction at a comparative level efficiency and transgenesis. The study showed the feasibility to increase the number of survived eggs and the capability to develop to blastocyst stage if gene concentration reduced to 2 pl/ml instead 4 pcl/ml, however both as gene and TE buffer microinjection are being led to less surviving in culture. The higher embryo implantation of the injected zygotes been noticed at lower gene concentration also (p<0.05). We demonstrated that the percentage of implantated embryos at two different gene concentrations is elevated to compare to those that generated blastocysts in a Petri dish: 36.7 % and 30 % in vivo after embryo transfer versus 8.6 % and 5.7 % in vitro culture. Due to in vitro culture is leading to unstable transgene expression, the embryo transfer directly after gene microinjection is preferable. The method described here can provide a strong basis for transgenesis efficiency 6.7 % and 3.3 % for two different gene concentrations 2 and 4 pl/ml, accordingly. We also demonstrated that 3 transgenic offspring from 20 newborn pups been identified successfully with PCR amplification of tail tissue samples.Read More