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Science ; 350(6264): 1101-4, 2015 Nov 27.
Article in English | MEDLINE | ID: mdl-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.


Subject(s)
Endogenous Retroviruses/genetics , Gene Targeting/methods , Retroviridae Infections/prevention & control , Swine/virology , Transplantation, Heterologous/methods , Virus Inactivation , Animals , Base Sequence , CRISPR-Cas Systems , Cell Line , Epithelial Cells/virology , Gene Dosage , Genes, pol , HEK293 Cells , Humans , Kidney/virology , Molecular Sequence Data , Retroviridae Infections/transmission , Retroviridae Infections/virology
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