利用向导RNA和Cas9核酸酶培育I类主要组织相容性复合体缺失的猪。
Creating class I MHC-null pigs using guide RNA and the Cas9 endonuclease.
作者信息
Reyes Luz M, Estrada Jose L, Wang Zheng Yu, Blosser Rachel J, Smith Rashod F, Sidner Richard A, Paris Leela L, Blankenship Ross L, Ray Caitlin N, Miner Aaron C, Tector Matthew, Tector A Joseph
机构信息
Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
Indiana University Health, Indianapolis, IN 46202; and
出版信息
J Immunol. 2014 Dec 1;193(11):5751-7. doi: 10.4049/jimmunol.1402059. Epub 2014 Oct 22.
Pigs are emerging as important large animal models for biomedical research, and they may represent a source of organs for xenotransplantation. The MHC is pivotal to the function of the immune system in health and disease, and it is particularly important in infection and transplant rejection. Pigs deficient in class I MHC could serve as important reagents to study viral immunity as well as allograft and xenograft rejection. In this study, we report the creation and characterization of class I MHC knockout pigs using the Cas9 nuclease and guide RNAs. Pig fetal fibroblasts were genetically engineered using Cas9 and guide RNAs, and class I MHC(-) cells were then used as nuclear donors for somatic cell nuclear transfer. We produced three piglets devoid of all cell surface class I proteins. Although these animals have reduced levels of CD4(-)CD8(+) T cells in peripheral blood, the pigs appear healthy and are developing normally. These pigs are a promising reagent for immunological research.
猪正成为生物医学研究中重要的大型动物模型，并且它们可能是异种移植器官的来源。主要组织相容性复合体（MHC）对于免疫系统在健康和疾病中的功能至关重要，在感染和移植排斥反应中尤为重要。缺乏I类MHC的猪可作为研究病毒免疫以及同种异体移植和异种移植排斥反应的重要试剂。在本研究中，我们报告了使用Cas9核酸酶和引导RNA创建和表征I类MHC基因敲除猪的过程。利用Cas9和引导RNA对猪胎儿成纤维细胞进行基因工程改造，然后将I类MHC（-）细胞用作体细胞核移植的核供体。我们培育出了三只缺乏所有细胞表面I类蛋白的仔猪。尽管这些动物外周血中CD4（-）CD8（+）T细胞水平降低，但这些猪看起来健康且发育正常。这些猪是免疫研究中很有前景的试剂。
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