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Modifying the common marmoset monkey (Callithrix jacchus) genome: transgenesis and targeted gene modification in vivo and in vitro

dc.contributor.advisorBehr, Rüdiger Prof. Dr.
dc.contributor.authorKahland, Tobias Sören
dc.titleModifying the common marmoset monkey (Callithrix jacchus) genome: transgenesis and targeted gene modification in vivo and in vitrode
dc.contributor.refereeWalter, Lutz Prof. Dr.
dc.description.abstractengThe most widely used animal model for biomedical research is the mouse. Yet, the physiological differences between mice and humans sometimes result in findings of questionable translational value. The common marmoset monkey (Callithrix jacchus) is a non-human primate, which is gaining more and more interest as a translational model organism. However, methods for the genetic modification, which have been well-established in the mouse for decades, have only recently become available for the common marmoset. In this study I applied different tools for transgenesis and targeted gene modification using the common marmoset as a model organism. The initial part of the study was performed at the Central Institute for Experimental Animals in Kawasaki, Japan, where I was successful in generating two transgenic common marmoset monkeys using the Early Transposon promoter and Oct4 and Sox2 enhancers (EOS) lentiviral vector. The EOS vector mediates pluripotencyassociated expression of enhanced Green Fluorescent Protein (eGFP) and, once introduced as a transgene, can be utilized as a fluorescence marker of pluripotent cells. The other parts of my work were accomplished at the German Primate Center in Goettingen. To test EOS activity in the common marmoset, fibroblasts of the EOS transgenic monkeys were reprogrammed to induced pluripotent stem (iPS) cells by non-viral means using the piggyBac transposon system. Notably, I was able to reprogram primary fibroblasts making use of only the four classical reprogramming factors SOX2, OCT4, KLF4 and c-MYC, omitting LIN28, which was reported to be essential for common marmoset monkey cell reprogramming. In the end I used a recent approach for targeted gene modification, the CRISPR/Cas9 system, to modify the Parkinson’s disease associated gene LRRK2 in common marmoset primary fibroblasts. In addition to that, I generated immortalized common marmoset monkey fibroblast cell lines by stably introducing the human telomerase reverse transcriptase (hTERT) transgene again using the piggyBac transposon. In conclusion, I successfully applied different strategies for the genetic modification of genes in the common marmoset genome. My study will hopefully contribute to an expanded application of the common marmoset in biomedical and translational
dc.contributor.coRefereePöhlmann, Stefan Prof. Dr.
dc.subject.engCommon marmosetde
dc.subject.engTargeted gene modificationde
dc.subject.engIn vitro fertilizationde
dc.subject.engiPS cellsde
dc.affiliation.instituteBiologische Fakultät für Biologie und Psychologiede
dc.subject.gokfullBiologie (PPN619462639)de

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