Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

Takehito Sugasawa
,
Takuro Nakano
,
Shin-ichiro Fujita
ORCID logo ,
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
ORCID logo ,
Yasushi Kawakami
,
Kazuhiro Takekoshi
*
Version 1 : Received: 1 July 2021 / Approved: 1 July 2021 / Online: 1 July 2021 (14:30:04 CEST)

A peer-reviewed article of this Preprint also exists.

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. 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.

Abstract

The World Anti-Doping Agency (WADA) has prohibited gene doping in the context of progress in gene therapy. In addition, there is a risk of the EPO gene being applied in gene doping among athletes. Along with this, development of a gene-doping test has been underway in worldwide. Here, we had two purposes: to develop a robust gene doping mouse model using the human EPO gene (hEPO) transferred using recombinant adenovirus (rAdV) as a vector and to develop a detection method to prove gene doping using this model. The rAdV including the hEPO gene were injected intravenously to transfer the gene to the liver. After injection, the mice developed significantly increased red blood cell counts in whole blood and increased gene expressions of hematopoietic markers in the spleen, indicating successful development of the gene doping model. Next, we detected direct and indirect proof of gene doping in whole blood DNA and RNA using qPCR assay and RNA sequencing. Proof was detected in one drop of whole blood DNA and RNA over a long period; furthermore, the overall RNA expression profiles significantly changed. Therefore, we have advanced detection of hEPO gene doping in humans.

Keywords

Gene doping; Gene therapy; Erythropoietin; Adenoviral vector; Sports; Athlete; RNA sequencing

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Download PDF Download Supplementary

Comments (0)

We encourage comments and feedback from a broad range of readers. See criteria for comments and our Diversity statement.

Leave a public comment
Send a private comment to the author(s)
* All users must log in before leaving a comment
Views 0
Downloads 0
Comments 0
Metrics 0
Get PDF Supplementary Files Cite Share 0
Bookmark BibSonomy Mendeley Reddit Delicious
×
Alerts
Notify me about updates to this article or when a peer-reviewed version is published.
We use cookies on our website to ensure you get the best experience.
Read more about our cookies here.