Detection Method for Gene Doping in a Mouse Model Expressing Human Erythropoietin from Adeno-Associated Virus Vector-9

• Published in: Genes Vol. 15; no. 6; p. 709
• Main Authors: Sugasawa, Takehito, Hirokawa, Atsushi, Otani, Norihiro, Kanki, Yasuharu, Nguyen, Kieu Dm, Takemasa, Tohru, Watanabe, Koichi, Takeuchi, Yoshinori, Yahagi, Naoya, Takahashi, Yoichiro
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 29.05.2024; MDPI
• Subjects: Animal models; Animals; Athletes; Blood; Coronaviruses; COVID-19; Dependovirus - genetics; DNA; DNA probes; Doping in Sports - methods; Drugs & sports; Enzymes; Erythropoietin; Erythropoietin - genetics; Ethylenediaminetetraacetic acid; Experimental methods; Experiments; Gene therapy; Genes; Genetic Therapy - methods; Genetic vectors; Genetic Vectors - genetics; Humans; Instrument industry; International economic relations; Male; Methods; Mice; Models, Animal; Research methodology; RNA; RNA probes; RNA sequencing; Canada; United States; Japan; gene doping; RNA-seq; mouse model; TaqMan-qPCR assay; erythropoietin
• ISSN: 2073-4425; 2073-4425
• DOI: 10.3390/genes15060709

With the rapid development of gene therapy technology in recent years, its abuse as a method of sports doping in athletics has become a concern. However, there is still room for improvement in gene-doping testing methods, and a robust animal model needs to be developed. Therefore, the purposes of this study were to establish a model of gene doping using recombinant adeno-associated virus vector-9, including the human erythropoietin gene (rAAV9-h ), and to establish a relevant testing method. First, it was attempted to establish the model using rAAV9-h on mice. The results showed a significant increase in erythrocyte volume accompanied by an increase in spleen weight, confirming the validity of the model. Next, we attempted to detect proof of gene doping by targeting DNA and RNA. Direct proof of gene doping was detected using a TaqMan-qPCR assay with certain primers/probes. In addition, some indirect proof was identified in RNAs through the combination of a TB Green qPCR assay with RNA sequencing. Taken together, these results could provide the foundation for an effective test for gene doping in human athletes in the future.