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Article

Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector

1
Laboratory of Clinical Examination/Sports Medicine, Division of Clinical Medicine, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8577, Japan
2
Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8577, Japan
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Research and Development Section, Anicom Specialty Medical Institute Inc., 2-6-3 Chojamachi 5F, Yokohama 231-0033, Japan
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Japan Society for the Promotion of Science, Kojimachi Business Center Building, Kojimachi, Chiyoda-ku, Tokyo 102-0083, Japan
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Department of Experimental Pathology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8577, Japan
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Analyst-Accenture Technology, Intelligent Platform Services, Accenture Japan Ltd., Akasaka Intercity AIR, 1-8-1 Akasaka, Minato-ku, Tokyo 107-8672, Japan
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Tsukuba i-Laboratory LLP, 2-1-17 Amakubo, Tsukuba 305-0005, Japan
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Department of Medical Technology, Faculty of Health Sciences, Tsukuba International University, 6-20-1 Manabe, Tsuchiura 300-0051, Japan
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Faculty of Health and Sport Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8574, Japan
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Juan Del Coso
Genes 2021, 12(8), 1249; https://doi.org/10.3390/genes12081249
Received: 23 June 2021 / Revised: 5 August 2021 / Accepted: 9 August 2021 / Published: 16 August 2021
(This article belongs to the Section Genetic Diagnosis)
Despite the World Anti-Doping Agency (WADA) ban on gene doping in the context of advancements in gene therapy, the risk of EPO gene-based doping among athletes is still present. To address this and similar risks, gene-doping tests are being developed in doping control laboratories worldwide. In this regard, the present study was performed with two objectives: to develop a robust gene-doping mouse model with the human EPO gene (hEPO) transferred using recombinant adenovirus (rAdV) as a vector and to develop a detection method to identify gene doping by using this model. The rAdV including the hEPO gene was injected intravenously to transfer the gene to the liver. After injection, the mice showed significantly increased whole-blood red blood cell counts and increased expression of hematopoietic marker genes in the spleen, indicating successful development of the gene-doping model. Next, direct and potentially indirect proof of gene doping were evaluated in whole-blood DNA and RNA by using a quantitative PCR assay and RNA sequencing. Proof of doping could be detected in DNA and RNA samples from one drop of whole blood for approximately a month; furthermore, the overall RNA expression profiles showed significant changes, allowing advanced detection of hEPO gene doping. View Full-Text
Keywords: gene doping; gene therapy; erythropoietin; adenoviral vector; sports; athlete; RNA sequencing gene doping; gene therapy; erythropoietin; adenoviral vector; sports; athlete; RNA sequencing
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MDPI and ACS Style

Sugasawa, T.; Nakano, T.; Fujita, S.-i.; Matsumoto, Y.; Ishihara, G.; Aoki, K.; Yanazawa, K.; Ono, S.; Tamai, S.; Manevich, L.; Ueda, H.; Ishibashi, N.; Tamai, K.; Kanki, Y.; Yoshida, Y.; Watanabe, K.; Takemasa, T.; Kawakami, Y.; Takekoshi, K. Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector. Genes 2021, 12, 1249. https://doi.org/10.3390/genes12081249

AMA Style

Sugasawa T, Nakano T, Fujita S-i, Matsumoto Y, Ishihara G, Aoki K, Yanazawa K, Ono S, Tamai S, Manevich L, Ueda H, Ishibashi N, Tamai K, Kanki Y, Yoshida Y, Watanabe K, Takemasa T, Kawakami Y, Takekoshi K. Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector. Genes. 2021; 12(8):1249. https://doi.org/10.3390/genes12081249

Chicago/Turabian Style

Sugasawa, Takehito, Takuro Nakano, Shin-ichiro Fujita, Yuki Matsumoto, Genki Ishihara, Kai Aoki, Koki Yanazawa, Seiko Ono, Shinsuke Tamai, Lev Manevich, Haruna Ueda, Noriyo Ishibashi, Kenshirou Tamai, Yasuharu Kanki, Yasuko Yoshida, Koichi Watanabe, Tohru Takemasa, Yasushi Kawakami, and Kazuhiro Takekoshi. 2021. "Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector" Genes 12, no. 8: 1249. https://doi.org/10.3390/genes12081249

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