최근 급속하게 발전하고 있는 유전자교정기술은 식물이 생산하는 특정 이차 대사산물의 집적을 유도하기 위한 식물대사공학연구에 아주 유용하게 이용되고 있다. 특히 이들 기술 들을 이용하여 만든 유전자교정 작물 중에 일부는 외래 DNA 단편이 잔존 하지 않기 때문에 기존의 유전자변형작물의 안전 관리규정에 적용되지 않을 수 있다는 장점이 있다. 따라서 본 리뷰는 phenylpropanoid 대사과정에 의하여 합성되는 다양한 종류의 이차대사산물을 집적시키기 위한 유전자교정 기술의 적용 연구결과 들을 조사하였다. 먼저, phenylpropanoid 생합성 대사과정에 관여하는 다양한 효소를 암호하는 유전자들을 목표로 하여 식물의 종에 따라 특이하게 집적되는 flavonoids, anthocyanin, 수용성 tannins, 로즈마린산 등의 집적을 유도하거나 화색을 변경하는 등의 성공적인 연구결과들을 검토하였다. 또한, phenylpropanoid 대사과정의 조절에 최종조절 스위치 역할을 하는 수많은 종류의 MYB 전사인자를 암호 하는 유전자를 목표로 하여 CRISPR 유전자교정을 시도한 연구결과들로부터 식물의 이차세포벽 형성에 관여하는 lignin, 물관부, cellulose 등의 생합성 조절 기작을 이해하고 MYB family에 속하는 수많은 종류의 유전자에 대한 개별적인 기능 분석 연구결과들을 조사 분석하여, 문제점 및 향후 연구 방향 등을 검토하였다.
Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at 37°C. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.
Animal experiments need numerous kinds of animal which are suitable for every research. About 300 mouse strains are developed up to the present, but they do not give satisfaction to every researchers. So we must build up the methods of breading animals which are newly developed and of maintenance of characteristics which were developed before. To maintain experimental animal is not only proceeding the generation but also increasing the animal populations, it needs geneticai control. Genetic factors which influence to reproduction are very important to maintain colony. These factors include lethal gene, chromosomal abberation, sterility gene, etc.. With the recent development of transgenic technology, scientists now can deliberately creat numerous specific animal models. To know how to manage the colony which has genetic defect on reproduction and transgenic mice is one of the key to study in vitro fertilization.