The embryonic genome activation (EGA) is genetically activated states that embryos make the materials such as growth factors for using themselves. EGA is various because they have many materials, different site, different stage, also different species. At this time, transcription factors are expressed. Transcription factors bind to specific DNA region, and regulate the gene expression. Thus, we check the expression of transcription factors, we can know that embryo development is very well or not. The development stages of embryos are basically the stages from fertilization to blastocyst. So, we check the embryos oocyte to blastocyst. In our experiments, we focus the early developmental transcription factors such as Cdx2, Oct4, Sox2, Nanog and E-Cadherin. Above antibody factors showed different expression sites, and there were many differentiated parts from other animal species. In addition, we compared the SCNT and parthenogenetic activation (PA) because these are same methods using electrical activation among the embryo production methods. Our results showed not only similar patterns but also different patterns between pig and mouse. Therefore, we have to investigate that different patterns of transcription factors play a role in pigs, and why occur.

The early-onset familial Alzheimer's disease (EOFAD/ FAD), the less common type of Alzheimer's disease (AD) currently affects a vast number of individuals worldwide. This type is being inherited as an autosomal dominant fashion. Missense mutations on Amyloid precursor protein (APP) and Presenilins 1 and 2 (PSEN1 & PSEN2) are known as major genetic factors in FAD. Conversely, missense mutations on microtubule-associated protein tau (MAPT) are also thought to involve. Up to date, several triple-transgenic animal models with muted forms of the human APP, PSENs and MAPT have been reported. Compared to other animals, canines are more emotional and their disease signs can be easily diagnosed. This attempt was to develop a triple transgenic canine model for the AD. We have obtained the coding sequences of APP, PSEN1 and MAPT from Dana-Farber/Harvard Cancer Center DNA resource core at HMS and incorporated several common AD mutations. The transgenic construct is composed of hNSE (ENO2) promoter-driven three AD genes fused together with modified 2A sequences. It was transfected into the canine fetal fibroblasts which were then used to perform somatic cell nuclear transfer (SCNT). The viable transgenic embryos were obtained after in vitro culture and the GFP was detected. In this study, we have successfully produced viable triple transgenic canine cloned embryos using SCNT technique. These transgenic canine embryos will be further developed into canines with FAD. The transgenic canines will be a good candidate in the AD research field.

Until now, problems related to shortage of organ for transplantation have been continuing. Pigs are the most suitable animal for xenotransplantation. Although primates are most similar to humans, they are not suitable because they have low productivity. Pigs are more productive than primates, and their organ size and physiological characteristics are similar to humans, with the exception of primates. In this study, we breeding the transgenic minipigs using natural mating to produce transgenic pigs. And, transgenic pigs has transmission rate that follow mendel’s rule. There are 20% hDAF gene, 20% US11 gene and 50% both hDAF and US11 gene in transgenic offsprings. Furthermore, transgenic pigs followed normal litter size, and piglets also has normal sex ratio. To suppress the immune function, experiments were performed using porcine ear fibroblast that transfected with hDAF and US11gene. In Cytotoxicity experiment against human complement, hDAF gene and double transgenic cell with both hDAF and US11 gene showed effect to reduce cytotoxicity rate in all of human complement condition. US11 gene and double transgenic cell were significantly reduce the cytotoxicity ratio in human NK cell. Besides, hDAF gene transgenic cell also reduce immune response in 10:1 concentration of human NK cell. In conclusion, natural mating was efficient method for breeding transgenic pigs. And, hDAF and US11 genes has effect for reduce cytotoxicity against human NK cell and human complement conditions.