Cas9-expressing chickens and pigs as resources for genome editing in livestock.


Journal

Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876

Informations de publication

Date de publication:
09 03 2021
Historique:
entrez: 4 3 2021
pubmed: 5 3 2021
medline: 17 8 2021
Statut: ppublish

Résumé

Genetically modified animals continue to provide important insights into the molecular basis of health and disease. Research has focused mostly on genetically modified mice, although other species like pigs resemble the human physiology more closely. In addition, cross-species comparisons with phylogenetically distant species such as chickens provide powerful insights into fundamental biological and biomedical processes. One of the most versatile genetic methods applicable across species is CRISPR-Cas9. Here, we report the generation of transgenic chickens and pigs that constitutively express Cas9 in all organs. These animals are healthy and fertile. Functionality of Cas9 was confirmed in both species for a number of different target genes, for a variety of cell types and in vivo by targeted gene disruption in lymphocytes and the developing brain, and by precise excision of a 12.7-kb DNA fragment in the heart. The Cas9 transgenic animals will provide a powerful resource for in vivo genome editing for both agricultural and translational biomedical research, and will facilitate reverse genetics as well as cross-species comparisons.

Identifiants

pubmed: 33658378
pii: 2022562118
doi: 10.1073/pnas.2022562118
pmc: PMC7958376
pii:
doi:

Types de publication

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

Langues

eng

Sous-ensembles de citation

IM

Informations de copyright

Copyright © 2021 the Author(s). Published by PNAS.

Déclaration de conflit d'intérêts

Competing interest statement: L.C. holds a patent on gene therapy vectors for treating cardiomyopathy that was licensed to DiNAQOR, is a member of the Scientific Advisory Board, and has shares in DiNAQOR.

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Auteurs

Beate Rieblinger (B)

Livestock Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Hicham Sid (H)

Reproductive Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Denise Duda (D)

Reproductive Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Tarik Bozoglu (T)

Clinic and Polyclinic for Internal Medicine I, Klinikum rechts der Isar, Technical University Munich, 81675 Munich, Germany.
Munich Heart Alliance, German Center for Cardiovascular Research, 81675 Munich, Germany.

Romina Klinger (R)

Reproductive Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Antonina Schlickenrieder (A)

Reproductive Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Kamila Lengyel (K)

Reproductive Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Krzysztof Flisikowski (K)

Livestock Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Tatiana Flisikowska (T)

Livestock Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Nina Simm (N)

Livestock Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Alessandro Grodziecki (A)

Livestock Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Carolin Perleberg (C)

Livestock Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Andrea Bähr (A)

Clinic and Polyclinic for Internal Medicine I, Klinikum rechts der Isar, Technical University Munich, 81675 Munich, Germany.
Munich Heart Alliance, German Center for Cardiovascular Research, 81675 Munich, Germany.

Lucie Carrier (L)

Institute of Experimental Pharmacology and Toxicology, University Medical Center Hamburg Eppendorf, 20246 Hamburg, Germany.
Institute of Experimental Pharmacology and Toxicology, German Centre for Cardiovascular Research, 20246 Hamburg, Germany.

Mayuko Kurome (M)

Gene Center, Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
Molecular Animal Breeding and Biotechnology, Department of Veterinary Sciences, Ludwig-Maximilians-Universität München, 81377 München, Germany.

Valeri Zakhartchenko (V)

Gene Center, Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
Molecular Animal Breeding and Biotechnology, Department of Veterinary Sciences, Ludwig-Maximilians-Universität München, 81377 München, Germany.

Barbara Kessler (B)

Gene Center, Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
Molecular Animal Breeding and Biotechnology, Department of Veterinary Sciences, Ludwig-Maximilians-Universität München, 81377 München, Germany.

Eckhard Wolf (E)

Gene Center, Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
Molecular Animal Breeding and Biotechnology, Department of Veterinary Sciences, Ludwig-Maximilians-Universität München, 81377 München, Germany.

Lutz Kettler (L)

Zoology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Harald Luksch (H)

Zoology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany.

Ibrahim T Hagag (IT)

Department of Veterinary Medicine, Institute of Virology, Freie Universität Berlin, 14163 Berlin, Germany.

Daniel Wise (D)

Department of Pathology, University of Cambridge, CB2 1QP Cambridge, United Kingdom.

Jim Kaufman (J)

Department of Pathology, University of Cambridge, CB2 1QP Cambridge, United Kingdom.
Institute for Immunology and Infection Research, School of Biological Sciences, University of Edinburgh, EH9 3FL Edinburgh, United Kingdom.

Benedikt B Kaufer (BB)

Department of Veterinary Medicine, Institute of Virology, Freie Universität Berlin, 14163 Berlin, Germany; b.kaufer@fu-berlin.de christian.kupatt@tum.de angelika.schnieke@wzw.tum.de benjamin.schusser@tum.de.

Christian Kupatt (C)

Clinic and Polyclinic for Internal Medicine I, Klinikum rechts der Isar, Technical University Munich, 81675 Munich, Germany; b.kaufer@fu-berlin.de christian.kupatt@tum.de angelika.schnieke@wzw.tum.de benjamin.schusser@tum.de.
Munich Heart Alliance, German Center for Cardiovascular Research, 81675 Munich, Germany.

Angelika Schnieke (A)

Livestock Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany; b.kaufer@fu-berlin.de christian.kupatt@tum.de angelika.schnieke@wzw.tum.de benjamin.schusser@tum.de.

Benjamin Schusser (B)

Reproductive Biotechnology, Department of Molecular Life Sciences, School of Life Sciences Weihenstephan, Technical University Munich, 85354 Freising, Germany; b.kaufer@fu-berlin.de christian.kupatt@tum.de angelika.schnieke@wzw.tum.de benjamin.schusser@tum.de.

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