{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T01:52:32Z","timestamp":1791597152007,"version":"4.3.4"},"reference-count":24,"publisher":"PeerJ","license":[{"start":{"date-parts":[[2021,10,20]],"date-time":"2021-10-20T00:00:00Z","timestamp":1634688000000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science, and Technology, Japan","doi-asserted-by":"crossref","award":["20H04062"],"award-info":[{"award-number":["20H04062"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Japan Sports Agency"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Gene doping is the misuse of genome editing and gene therapy technologies for the purpose of manipulating specific genes or gene functions in order to improve athletic performance. However, a non-invasive detection method for gene doping using recombinant adenoviral (rAdV) vectors containing human follistatin (\n                      <jats:italic>hFST<\/jats:italic>\n                      ) genes (rAdV&lt;\n                      <jats:italic>hFST<\/jats:italic>\n                      &gt;) has not yet been developed. Therefore, the aim of this study was to develop a method to detect gene doping using rAdV&lt;\n                      <jats:italic>hFST<\/jats:italic>\n                      &gt;.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      First, we generated rAdV&lt;\n                      <jats:italic>hFST<\/jats:italic>\n                      &gt; and evaluated the overexpression of the\n                      <jats:italic>hFST<\/jats:italic>\n                      gene, FST protein, and muscle protein synthesis signaling using cell lines. Next, rAdV&lt;\n                      <jats:italic>hFST<\/jats:italic>\n                      &gt; was injected intravenously or intramuscularly into mice, and whole blood was collected, and\n                      <jats:italic>hFST<\/jats:italic>\n                      and cytomegalovirus promoter (\n                      <jats:italic>CMVp<\/jats:italic>\n                      ) gene fragments were detected using TaqMan-quantitative polymerase chain reaction (qPCR). Finally, to confirm the specificity of the primers and the TaqMan probes, samples from each experiment were pooled, amplified using TaqMan-qPCR, and sequenced using the Sanger sequencing.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The expression of hFST and FST proteins and muscle protein synthesis signaling significantly increased in C2C12 cells. In long-term, transgene fragments could be detected until 4 days after intravenous injection and 3 days after intramuscular injection. Finally, the Sanger sequencing confirmed that the primers and TaqMan probe specifically amplified the gene sequence of interest.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      These results indicate the possibility of detecting gene doping using rAdV&lt;\n                      <jats:italic>hFST<\/jats:italic>\n                      &gt; using TaqMan-qPCR in blood samples. This study may contribute to the development of detection methods for gene doping using rAdV&lt;\n                      <jats:italic>hFST<\/jats:italic>\n                      &gt;.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.7717\/peerj.12285","type":"journal-article","created":{"date-parts":[[2021,10,20]],"date-time":"2021-10-20T08:23:19Z","timestamp":1634718199000},"page":"e12285","source":"Crossref","is-referenced-by-count":12,"title":["Development of a gene doping detection method to detect overexpressed human follistatin using an adenovirus vector in mice"],"prefix":"10.7717","volume":"9","author":[{"given":"Koki","family":"Yanazawa","sequence":"first","affiliation":[{"name":"Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takehito","family":"Sugasawa","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Examination\/Sports Medicine, Division of Clinical Medicines, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Aoki","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Examination\/Sports Medicine, Division of Clinical Medicines, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takuro","family":"Nakano","sequence":"additional","affiliation":[{"name":"Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasushi","family":"Kawakami","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Examination\/Sports Medicine, Division of Clinical Medicines, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazuhiro","family":"Takekoshi","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Examination\/Sports Medicine, Division of Clinical Medicines, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4443","published-online":{"date-parts":[[2021,10,20]]},"reference":[{"key":"10.7717\/peerj.12285\/ref-1","doi-asserted-by":"publisher","first-page":"e8595","DOI":"10.7717\/peerj.8595","article-title":"The detection of trans gene fragments of hEPO in gene doping model mice by TaqMan qPCR assay","volume":"8","author":"Aoki","year":"2020","journal-title":"PeerJ"},{"issue":"11","key":"10.7717\/peerj.12285\/ref-2","doi-asserted-by":"publisher","first-page":"1014","DOI":"10.1038\/ncb1101-1014","article-title":"Akt\/mTOR pathway is a crucial regulator of skeletal muscle hypertrophy and can prevent muscle atrophy in vivo","volume":"3","author":"Bodine","year":"2001","journal-title":"Nature Cell Biology"},{"issue":"4","key":"10.7717\/peerj.12285\/ref-3","doi-asserted-by":"publisher","first-page":"328","DOI":"10.3181\/00379727-214-44100","article-title":"Inhibins, activins, and follistatins: the saga continues","volume":"214","author":"DePaolo","year":"1997","journal-title":"Proceedings of the Society for Experimental Biology & Medicine. 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