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Article

Generation and Breeding of EGFP-Transgenic Marmoset Monkeys: Cell Chimerism and Implications for Disease Modeling

by
Charis Drummer
1,2,*,
Edgar-John Vogt
1,
Michael Heistermann
3,
Berit Roshani
4,
Tamara Becker
5,
Kerstin Mätz-Rensing
6,
Wilfried A. Kues
7,
Sebastian Kügler
8 and
Rüdiger Behr
1,2,*
1
Platform Degenerative Diseases, German Primate Center–Leibniz Institute for Primate Research, Kellnerweg 4, 37077 Göttingen, Germany
2
DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, 37099 Göttingen, Germany
3
Endocrinology Laboratory, German Primate Center–Leibniz Institute for Primate Research, Kellnerweg 4, 37077 Göttingen, Germany
4
Unit of Infection Models, German Primate Center–Leibniz Institute for Primate Research, Kellnerweg 4, 37077 Göttingen, Germany
5
Primate Husbandry, German Primate Center–Leibniz Institute for Primate Research, Kellnerweg 4, 37077 Göttingen, Germany
6
Pathology Unit, German Primate Center–Leibniz-Institute for Primate Research, Kellnerweg 4, 37077 Göttingen, Germany
7
Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Mariensee, 31535 Neustadt, Germany
8
Center for Nanoscale Microscopy and Physiology of the Brain (CNMPB) at Department of Neurology, University of Göttingen, Waldweg 33, 37073 Göttingen, Germany
*
Authors to whom correspondence should be addressed.
Cells 2021, 10(3), 505; https://doi.org/10.3390/cells10030505
Submission received: 4 February 2021 / Revised: 18 February 2021 / Accepted: 22 February 2021 / Published: 27 February 2021
(This article belongs to the Section Stem Cells)
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Abstract

Genetic modification of non-human primates (NHP) paves the way for realistic disease models. The common marmoset is a NHP species increasingly used in biomedical research. Despite the invention of RNA-guided nucleases, one strategy for protein overexpression in NHP is still lentiviral transduction. We generated three male and one female enhanced green fluorescent protein (EGFP)-transgenic founder marmosets via lentiviral transduction of natural preimplantation embryos. All founders accomplished germline transmission of the transgene by natural mating, yielding 20 transgenic offspring together (in total, 45 pups; 44% transgenic). This demonstrates that the transgenic gametes are capable of natural fertilization even when in competition with wildtype gametes. Importantly, 90% of the transgenic offspring showed transgene silencing, which is in sharp contrast to rodents, where the identical transgene facilitated robust EGFP expression. Furthermore, we consistently discovered somatic, but so far, no germ cell chimerism in mixed wildtype/transgenic litters. Somatic cell chimerism resulted in false-positive genotyping of the respective wildtype littermates. For the discrimination of transgenic from transgene-chimeric animals by polymerase chain reaction on skin samples, a chimeric cell depletion protocol was established. In summary, it is possible to establish a cohort of genetically modified marmosets by natural mating, but specific requirements including careful promoter selection are essential.
Keywords: marmoset monkey; non-human primate; embryo; genetic modification; germline transmission; germ cell; transgenesis; chimerism; hematopoietic stem cell marmoset monkey; non-human primate; embryo; genetic modification; germline transmission; germ cell; transgenesis; chimerism; hematopoietic stem cell

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MDPI and ACS Style

Drummer, C.; Vogt, E.-J.; Heistermann, M.; Roshani, B.; Becker, T.; Mätz-Rensing, K.; Kues, W.A.; Kügler, S.; Behr, R. Generation and Breeding of EGFP-Transgenic Marmoset Monkeys: Cell Chimerism and Implications for Disease Modeling. Cells 2021, 10, 505. https://doi.org/10.3390/cells10030505

AMA Style

Drummer C, Vogt E-J, Heistermann M, Roshani B, Becker T, Mätz-Rensing K, Kues WA, Kügler S, Behr R. Generation and Breeding of EGFP-Transgenic Marmoset Monkeys: Cell Chimerism and Implications for Disease Modeling. Cells. 2021; 10(3):505. https://doi.org/10.3390/cells10030505

Chicago/Turabian Style

Drummer, Charis, Edgar-John Vogt, Michael Heistermann, Berit Roshani, Tamara Becker, Kerstin Mätz-Rensing, Wilfried A. Kues, Sebastian Kügler, and Rüdiger Behr. 2021. "Generation and Breeding of EGFP-Transgenic Marmoset Monkeys: Cell Chimerism and Implications for Disease Modeling" Cells 10, no. 3: 505. https://doi.org/10.3390/cells10030505

APA Style

Drummer, C., Vogt, E.-J., Heistermann, M., Roshani, B., Becker, T., Mätz-Rensing, K., Kues, W. A., Kügler, S., & Behr, R. (2021). Generation and Breeding of EGFP-Transgenic Marmoset Monkeys: Cell Chimerism and Implications for Disease Modeling. Cells, 10(3), 505. https://doi.org/10.3390/cells10030505

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