論文 - 詳細
| RRC ID | 69027 |
|---|---|
| 著者 | Yanazawa K, Sugasawa T, Aoki K, Nakano T, Kawakami Y, Takekoshi K. |
| タイトル | Development of a gene doping detection method to detect overexpressed human follistatin using an adenovirus vector in mice. |
| ジャーナル | PeerJ |
| Abstract |
Background: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 (hFST) genes (rAdV<hFST>) has not yet been developed. Therefore, the aim of this study was to develop a method to detect gene doping using rAdV<hFST>. Methods:First, we generated rAdV<hFST> and evaluated the overexpression of the hFST gene, FST protein, and muscle protein synthesis signaling using cell lines. Next, rAdV<hFST> was injected intravenously or intramuscularly into mice, and whole blood was collected, and hFST and cytomegalovirus promoter (CMVp) 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. Results: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. Conclusions:These results indicate the possibility of detecting gene doping using rAdV<hFST> using TaqMan-qPCR in blood samples. This study may contribute to the development of detection methods for gene doping using rAdV<hFST>. |
| 巻・号 | 9 |
| ページ | e12285 |
| 公開日 | 2021-1-1 |
| DOI | 10.7717/peerj.12285 |
| PII | 12285 |
| PMID | 34721976 |
| PMC | PMC8541302 |
| IF | 2.379 |
| リソース情報 | |
| ヒト・動物細胞 | HuH-7(RCB1366) C2C12(RCB0987) |