The Axolotl as a system to define the function and evolution of reprogramming activities
Zellbiologie
Zusammenfassung der Projektergebnisse
In summary, we have shown that the duplication of ClassV Pou genes - OCT4 and Pou2 - once thought to have occured at the base of the mammalian lineage actually occured atleast as early as the first tetrapodian lineage, as we identified clear orthologs to both paralogs in the axolotl. We further showed that these proteins have the ability to induce a bonafide pluripotent state in human fibroblasts. We further have developed gene and protein knockout/down approaches including CRISPR mediated gene deletion to show that Sox2 and Oct4 play central roles in the amplification of neural stem cells during axolotl spinal cord regeneration. Finally we have developed the transgenic reporter animals, as well as transplantation assays to functionally assay if Oct4 and Sox2 can are involved in conferring a potential pluripotent character to these neural stem cells if provided a permissive environment.
Projektbezogene Publikationen (Auswahl)
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(2012) Regeneration and reprogramming. Curr Opin Genet Dev. 22:485-93
Knapp D, Tanaka EM
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(2012) Reprogramming to pluripotency is an ancient trait of vertebrate Oct4 and Pou2 proteins. Nature Communications, 3:1279
Tapia N, Reinhardt P,Duemmler A, Wu G, Araúzo-Bravo MJ, Esch D, Greber B, Cojocaru V, Rascon CA, Tazaki A, Kump K, Voss R, Tanaka EM, Schöler HR
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(2013). Germline transgenic methods for tracking cells and testing gene function during regeneration in the axolotl, Stem Cell Reports, 1:90-103
Khattak S, Schuez M, Richter T , Knapp D, Haigo SL, Sandoval-Guzmán T, Hradlikova K, Duemmler A, Kerney R, Tanaka EM
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(2014). CRISPR- mediated genomic deletion of Sox2 in the axolotl shows a requirement in spinal cord neural stem cell amplification during tail regeneration. Stem Cell Reports
Fei JF, Schuez M, Tazaki A, Taniguchi Y, Roensch K, Tanaka EM
