Dose Control and Effect Analysis of Transgenic Microinjection
Posted by Andrea Brook on December 24th, 2015
Transgenic technology is widely used in the field of life sciences in recent emerging technologies. At present, more than 80% of transgenic animal models are completed by microinjection. The exogenous gene microinjection in animal embryo is a key step. The injection mode and injection dose directly influence the survival rate of embryos in vitro development and the success rate of transgenic animals.
However, in the previous studies, the dose control of exogenous gene injection using hydraulic hand controlled by micro injector is not objective. It is often causes astnchronous development by injecting astnchronous dose in the same culture environment, which reduces the whole effect of exogenous genes.
In this study, the relationship among different injection pressure, different injection time and injection dose is studied by using a more advanced air pressure controlled micro injector, which can achieve the goal of constant injection dose and reduce the error of repeated injection. And the optimal injection dose is found through the analysis of the development effect of the embryos after injection. The effect of injection on the early development of the embryo is reduced; the integration rate of exogenous gene is improved; the good condition for the preparation of transgenic animal model is created.
In order to analyze the impact of transgenic microinjection of the injected dose effecting on embryonic development. We use a tip diameter of 0.5 ~ 1.0um microinjection needle microinjection pressure testing under different conditions and at different times, and injecting 133 fertilized mouse eggs with different doses. Then we observe the development situation after injection.
The pressure controlled micro injector used in the experiment has the characteristics of short injection time (msec), controllable controllability and good repeatability. Under the same conditions, the error is less than 0.004pl. The results shows that when the injected dose was 2 ~ 4PL, the embryo development is not affected. When the embryos are more than 10pl, all the embryos die 1h after injection. It is proved that the main factors that cause cell damage and influence the development of the embryo is the dosage when the injection needle tip diameter is less than 1.0um.