猪内源性逆转录病毒（PERVs）的全基因组灭活。

Genome-wide inactivation of porcine endogenous retroviruses (PERVs).

机构信息

Department of Genetics, Harvard Medical School, Boston, MA, USA. Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA. eGenesis Biosciences, Boston, MA 02115, USA.

Department of Genetics, Harvard Medical School, Boston, MA, USA. College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.

出版信息

Science. 2015 Nov 27;350(6264):1101-4. doi: 10.1126/science.aad1191. Epub 2015 Oct 11.

DOI:10.1126/science.aad1191

PMID:26456528

Abstract

The shortage of organs for transplantation is a major barrier to the treatment of organ failure. Although porcine organs are considered promising, their use has been checked by concerns about the transmission of porcine endogenous retroviruses (PERVs) to humans. Here we describe the eradication of all PERVs in a porcine kidney epithelial cell line (PK15). We first determined the PK15 PERV copy number to be 62. Using CRISPR-Cas9, we disrupted all copies of the PERV pol gene and demonstrated a >1000-fold reduction in PERV transmission to human cells, using our engineered cells. Our study shows that CRISPR-Cas9 multiplexability can be as high as 62 and demonstrates the possibility that PERVs can be inactivated for clinical application of porcine-to-human xenotransplantation.

摘要

器官短缺是器官衰竭治疗的主要障碍。虽然猪器官被认为很有前途，但人们对猪内源性逆转录病毒（PERVs）向人类传播的担忧限制了其使用。在这里，我们描述了在猪肾上皮细胞系（PK15）中消除所有 PERVs 的方法。我们首先确定 PK15 PERV 的拷贝数为 62。使用 CRISPR-Cas9，我们破坏了 PERV pol 基因的所有拷贝，并证明了用我们工程化的细胞，PERV 的传播到人类细胞的减少超过了 1000 倍。我们的研究表明，CRISPR-Cas9 的多重性可以高达 62，并证明了 PERVs 可以被失活，用于猪到人的异种移植的临床应用。