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

Genome-wide inactivation of porcine endogenous retroviruses (PERVs)

Luhan Yang  1 , Marc Güell  2 , Dong Niu  3 , Haydy George  4 , Emal Lesha  4 , Dennis Grishin  4 , John Aach  4 , Ellen Shrock  4 , Weihong Xu  5 , Jürgen Poci  4 , Rebeca Cortazio  4 , Robert A Wilkinson  6 , Jay A Fishman  6 , George Church  1
Affiliations

Genome-wide inactivation of porcine endogenous retroviruses (PERVs)

Luhan Yang et al. Science. .

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.

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