Prion(Pr) protein plays a pivotal role in the infection of bovine spongiform encephalopathy(BSE). The aim of the present research was to make Pr gene knockout(KO) cows,which lack Pr mRNA or protein,and to reveal their...
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Prion(Pr) protein plays a pivotal role in the infection of bovine spongiform encephalopathy(BSE). The aim of the present research was to make Pr gene knockout(KO) cows,which lack Pr mRNA or protein,and to reveal their characteristics for estimating the *** were successful in production of Pr gene homo KO Japanese Black cows using leading-edge genetic engineering techniques,cell culture techniques and animal reproductive techniques(somatic cell nuclear transfer:SCNT and so on) as follows:Firstly,hetero Pr gene was knockouted in fibroblastic cells derived from bovine *** somatic cell nucleus with hetero Pr gene KO was transferred into oocyte without germinal vesicle(GV).After activation and in vitro culture,somatic cell nucleus of the somatic cell nuclear cloning embryo at blastocyst stage was pulled out and transferred into oocyte without GV to make homo Pr gene KO *** homo Pr gene KO embryos were transferred in to the uterus. In the present meeting,we would like to show and discuss the procedure to make Pr gene homo KO cows and their usefulness,and to propose the importance of animal biotechnological techniques on food safety and public ***,detailed data of Pr distribution in organs of somatic cell nuclear cloning Japanese Black cows (used as controls) will be demonstrated as follows:A sandwich enzyme linked immunosorbent assay(ELISA) revealed that high levels of Pr proteinrotein(9.0-11.0μg/g wet tissue) was demonstrated in brain of somatic cell nuclear transfer-cows(wild-type control animals).Lower levels of Pr proteinrotein were shown in other organs as follows in decreasing order:skeletal muscles(longissimus,iliocostalis thoracis,splenius,femoris biceps,triceps brachii,longissimus thoracis and omotransversarius muscles),intestinal tracts(ileum,jejunum,duodenum,colon and cecum) and tongue(apex,posterior,anterior and radix areas)(approx.5.0-to 31.0-fold less Pr proteinrotein than the brain).Immunohistochemical analyses showed t
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