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Efficient production of biallelic GGTA1 knockout pigs by cytoplasmic microinjection of CRISPR/Cas9 into zygotes

Efficient production of biallelic GGTA1 knockout pigs by cytoplasmic microinjection of... Friedrich-Loeffler-Institut, Mariensee, Neustadt, Germany Background: Xenotransplantation is considered to be a promising solution to the growing demand for suitable donor organs for transplantation. Despite tremendous Correspondence Bjoern Petersen or Heiner Niemann, progress in the generation of pigs with multiple genetic modifications thought to be Institute of Farm Animal Genetics, necessary to overcoming the severe rejection responses after pig- to- non- human pri- Friedrich-Loeffler-Institut, Neustadt am Ruebenberge/Mariensee, Germany. mate xenotransplantation, the production of knockout pigs by somatic cell nuclear Emails: bjoern.petersen@fli.bund.de; transfer (SCNT) is still an inefficient process. Producing genetically modified pigs by heiner.niemann@fli.bund.de intracytoplasmic microinjection of porcine zygotes is an alluring alternative. The por- cine GGTA1 gene encodes for the α1,3- galactosyltransferase that synthesizes the Gal epitopes on porcine cells which constitute the major antigen in a xenotransplantation setting. GGTA1- KO pigs have successfully been produced by transfecting somatic cells with zinc- finger nucleases (ZFNs), transcription activator- like effector nucleases (TALENs), or CRISPR/Cas targeting GGTA1, followed by SCNT. Methods: Here, we microinjected a CRISPR/Cas9 vector coding for a single-guide RNA (sgRNA) targeting exon 8 of the GGTA1 gene into the cytoplasm of 97 in vivo-derived porcine zygotes and transferred 86 of the microinjected embryos into three hormonally synchronized recipients. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Xenotransplantation Wiley

Efficient production of biallelic GGTA1 knockout pigs by cytoplasmic microinjection of CRISPR/Cas9 into zygotes

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References (44)

Publisher
Wiley
Copyright
© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
ISSN
0908-665X
eISSN
1399-3089
DOI
10.1111/xen.12258
pmid
27610605
Publisher site
See Article on Publisher Site

Abstract

Friedrich-Loeffler-Institut, Mariensee, Neustadt, Germany Background: Xenotransplantation is considered to be a promising solution to the growing demand for suitable donor organs for transplantation. Despite tremendous Correspondence Bjoern Petersen or Heiner Niemann, progress in the generation of pigs with multiple genetic modifications thought to be Institute of Farm Animal Genetics, necessary to overcoming the severe rejection responses after pig- to- non- human pri- Friedrich-Loeffler-Institut, Neustadt am Ruebenberge/Mariensee, Germany. mate xenotransplantation, the production of knockout pigs by somatic cell nuclear Emails: bjoern.petersen@fli.bund.de; transfer (SCNT) is still an inefficient process. Producing genetically modified pigs by heiner.niemann@fli.bund.de intracytoplasmic microinjection of porcine zygotes is an alluring alternative. The por- cine GGTA1 gene encodes for the α1,3- galactosyltransferase that synthesizes the Gal epitopes on porcine cells which constitute the major antigen in a xenotransplantation setting. GGTA1- KO pigs have successfully been produced by transfecting somatic cells with zinc- finger nucleases (ZFNs), transcription activator- like effector nucleases (TALENs), or CRISPR/Cas targeting GGTA1, followed by SCNT. Methods: Here, we microinjected a CRISPR/Cas9 vector coding for a single-guide RNA (sgRNA) targeting exon 8 of the GGTA1 gene into the cytoplasm of 97 in vivo-derived porcine zygotes and transferred 86 of the microinjected embryos into three hormonally synchronized recipients.

Journal

XenotransplantationWiley

Published: Sep 1, 2016

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