earticle

영어

To investigate the efficient laboratory techniques for the injection of TALENs mRNA into porcine parthenogenesis and IVF embryos we had explores the injection time, volume and concentration, for the efficient blastocyst production has been identified within the experimental period. In experiment 1, to investigate injection time, compared 4 different time durations (1 hr, 2 hrs, 4 hrs & 5 hrs) after post activation of parthenogenetic embryos. There is no significant difference between four groups regarding the percentage of cleavages (55%). But comparatively higher percentage (20%) of blastocyst can be observed in the groups of 1hr and 2hrs injection after post activation. Even though observed higher blastocysts, only EGFP expressed blastocysts (2.2%) taken from the treatment of injection after 4 hrs of post activation. In experiment 2, compared two different concentration of EGFP injection in to embryos. 20 ng/μL and 50 ng/μL two concentrations were injected to the parthenogenetic embryos after 4 hrs of post activation and observed blastocysts after 7 days of incubation. There were significant difference between two treatments of concentrations considering percentages of cleavages and blastocysts. 50 ng/μL concentration injected embryos capable enough to cleaved up to 39% and produces blastocysts 10%, meanwhile 20 ng/ μL concentration illustrates 62% cleavage and 16% blastocysts percentages. Only 20 ng/μL injected embryos capable enough to express EGFP (3.1%) after 7 days of incubation. These results implies that 20 ng/μL concentration and injection 4hrs after post activation more effective injection conditions for microinjection.