Extensive germline genome engineering in pigs.

Animals CRISPR-Associated Protein 9 / genetics CRISPR-Cas Systems Cells, Cultured Galactosyltransferases / genetics Gene Knockout Techniques Genetic Engineering / methods Germ Cells / metabolism Mixed Function Oxygenases / genetics N-Acetylgalactosaminyltransferases / genetics Sus scrofa / genetics Transplantation, Heterologous

Journal

Nature biomedical engineering

ISSN: 2157-846X

Titre abrégé: Nat Biomed Eng

Pays: England

ID NLM: 101696896

Informations de publication

Date de publication:
02 2021

Historique:

received: 29 01 2020

accepted: 22 08 2020

pubmed: 23 9 2020

medline: 13 3 2021

entrez: 22 9 2020

Statut: ppublish

Résumé

The clinical applicability of porcine xenotransplantation-a long-investigated alternative to the scarce availability of human organs for patients with organ failure-is limited by molecular incompatibilities between the immune systems of pigs and humans as well as by the risk of transmitting porcine endogenous retroviruses (PERVs). We recently showed the production of pigs with genomically inactivated PERVs. Here, using a combination of CRISPR-Cas9 and transposon technologies, we show that pigs with all PERVs inactivated can also be genetically engineered to eliminate three xenoantigens and to express nine human transgenes that enhance the pigs' immunological compatibility and blood-coagulation compatibility with humans. The engineered pigs exhibit normal physiology, fertility and germline transmission of the 13 genes and 42 alleles edited. Using in vitro assays, we show that cells from the engineered pigs are resistant to human humoral rejection, cell-mediated damage and pathogenesis associated with dysregulated coagulation. The extensive genome engineering of pigs for greater compatibility with the human immune system may eventually enable safe and effective porcine xenotransplantation.

Identifiants

DOI: 10.1038/s41551-020-00613-9 PMID: 32958897

pubmed: 32958897

doi: 10.1038/s41551-020-00613-9

pii: 10.1038/s41551-020-00613-9

doi:

Substances chimiques

Mixed Function Oxygenases EC 1.-

CMPacetylneuraminate monooxygenase EC 1.14.18.2

Galactosyltransferases EC 2.4.1.-

N-Acetylgalactosaminyltransferases EC 2.4.1.-

alpha-1,3-galactosyltransferase 1, porcine EC 2.4.1.-

beta-1,4-N-acetyl-galactosaminyl transferase 2 EC 2.4.1.165

CRISPR-Associated Protein 9 EC 3.1.-

Banques de données

figshare

['10.6084/m9.figshare.12841418.v1']

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

Pagination

134-143

Commentaires et corrections

Type : CommentIn

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