scholarly journals Synthetic certified DNA reference material for analysis of human erythropoietin transgene and transcript in gene doping and gene therapy

Gene Therapy ◽  
2016 ◽  
Vol 23 (10) ◽  
pp. 708-717 ◽  
Author(s):  
A Baoutina ◽  
S Bhat ◽  
M Zheng ◽  
L Partis ◽  
M Dobeson ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Reference Material ◽  
Gene Doping ◽  
Human Erythropoietin

scholarly journals The detection of trans gene fragments of hEPO in gene doping model mice by Taqman qPCR assay

2019 ◽  
Author(s):  
Kai Aoki ◽  
Takehito Sugasawa ◽  
Kouki Yanazawa ◽  
Koichi Watanabe ◽  
Tohru Takemasa ◽  
...  
Keyword(s):  
Whole Blood ◽  
Plasmid Vector ◽  
Sybr Green ◽  
Gene Doping ◽  
Human Erythropoietin ◽  
World Anti Doping Agency ◽  
Total Dna ◽  
Stool Dna ◽  
Tail Tip ◽  
Taqman Qpcr

BACKGROUND. With the rapid progress of genetic engineering and gene therapy methods, the World Anti-Doping Agency has raised concerns regarding gene doping, which is prohibited in sports. However, there is no standard method available for detecting transgenes delivered by injection of naked plasmids. Here, we developed a detection method for detecting transgenes delivered by injection of naked plasmids in a mouse model that mimics gene doping. METHODS. Whole blood from the tail tip and one piece of stool were used as pre-samples of injection. Next, a plasmid vector containing the human erythropoietin (hEPO) gene was injected into mice through intravenous (IV), intraperitoneal (IP), or local muscular (IM) injection. At 1, 2, 3, 6, 12, 24, and 48 h after injection, approximately 50 μL whole blood was collected from the tail tip. One piece of stool was collected at 6, 12, 24, and 48 h. From each sample, total DNA was extracted and transgene fragments were analyzed by Taqman quantitative PCR (qPCR) and SYBR green qPCR. RESULTS. In whole blood DNA samples evaluated by Taqman qPCR, the transgene fragments were detected at all time points in the IP sample and at 1, 2, 3, 6, and 12 h in the IV and IM samples. In the stool-DNA samples, the transgene fragments were detected at 6, 12, 24, and 48 h in the IV and IM samples by Taqman qPCR. In the analysis by SYBR green qPCR, the transgene fragments were detected at some time point in both specimens; however, many non-specific amplicons were detected. CONCLUSIONS. These results indicate that transgene fragments evaluated after each injection method of naked plasmids were detected in whole-blood and stool DNA samples. These findings may facilitate the development of methods for detecting gene doping.

scholarly journals The detection of trans gene fragments of hEPO in gene doping model mice by Taqman qPCR assay

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8595 ◽  
Author(s):  
Kai Aoki ◽  
Takehito Sugasawa ◽  
Kouki Yanazawa ◽  
Koichi Watanabe ◽  
Tohru Takemasa ◽  
...  
Keyword(s):  
Whole Blood ◽  
Plasmid Vector ◽  
Sybr Green ◽  
Gene Doping ◽  
Human Erythropoietin ◽  
World Anti Doping Agency ◽  
Total Dna ◽  
Stool Dna ◽  
Tail Tip ◽  
Taqman Qpcr

Background With the rapid progress of genetic engineering and gene therapy methods, the World Anti-Doping Agency has raised concerns regarding gene doping, which is prohibited in sports. However, there is no standard method available for detecting transgenes delivered by injection of naked plasmids. Here, we developed a detection method for detecting transgenes delivered by injection of naked plasmids in a mouse model that mimics gene doping. Methods Whole blood from the tail tip and one piece of stool were used as pre-samples of injection. Next, a plasmid vector containing the human erythropoietin (hEPO) gene was injected into mice through intravenous (IV), intraperitoneal (IP), or local muscular (IM) injection. At 1, 2, 3, 6, 12, 24, and 48 h after injection, approximately 50 µL whole blood was collected from the tail tip. One piece of stool was collected at 6, 12, 24, and 48 h. From each sample, total DNA was extracted and transgene fragments were analyzed by Taqman quantitative PCR (qPCR) and SYBR green qPCR. Results In whole blood DNA samples evaluated by Taqman qPCR, the transgene fragments were detected at all time points in the IP sample and at 1, 2, 3, 6, and 12 h in the IV and IM samples. In the stool-DNA samples, the transgene fragments were detected at 6, 12, 24, and 48 h in the IV and IM samples by Taqman qPCR. In the analysis by SYBR green qPCR, the transgene fragments were detected at some time point in both specimens; however, many non-specific amplicons were detected. Conclusions These results indicate that transgene fragments evaluated after each injection method of naked plasmids were detected in whole-blood and stool DNA samples. These findings may facilitate the development of methods for detecting gene doping.

scholarly journals Gene therapy for renal anemia in mice with polycystic kidney using an adenovirus vector encoding the human erythropoietin gene

1999 ◽  
Vol 55 (4) ◽  
pp. 1234-1240 ◽  
Author(s):  
Shiwori Osada ◽  
Isao Ebihara ◽  
Yasuhiro Setoguchi ◽  
Hisahide Takahashi ◽  
Yasuhiko Tomino ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Adenovirus Vector ◽  
Renal Anemia ◽  
Polycystic Kidney ◽  
Human Erythropoietin

scholarly journals Gene doping and bioethics: legal regulation. Prospects, forecasts, obstacles

2021 ◽  
Vol 1 (8) ◽  
pp. 62-68
Author(s):  
O. A. Shevchenko
Keyword(s):  
Gene Therapy ◽  
National Level ◽  
Modern Science ◽  
Legal Regulation ◽  
Point Of View ◽  
Future Generations ◽  
Legal Research ◽  
Gene Doping ◽  
Ethical Point ◽  
Genetic Science

The article analyzes the correlation between the development of genetic science and bioethics issues. In the context of the rapid and steady development of genetic engineering and biomedicine, there is a lag in legal research in this area. Determining the vector of development of legal regulation of gene therapy and gene doping is currently one of the most important issues of modern science, which needs to be resolved from a legal and ethical point of view. In regulatory legal acts in the field of international sports and at the national level, a ban on the use of gene doping has been established, as well as responsibility for its use is provided. However, the measures taken are not enough.The article considers some existing problematic aspects of the correlation between the development of genetic science and bioethics and suggests ways to solve them. Thus, it is proposed to build a system of principles for preventing and countering the use of the gene doping method and to differentiate the concepts of gene therapy and gene doping in order to comply with the principles of Olympism and preserve the health of future generations.

Development of a dCas9-Based High-Throughput Gene Doping Analysis (HiGDA) for Human Erythropoietin

2021 ◽  
Author(s):  
Joon-Yeop Yi ◽  
Minyoung Kim ◽  
Byung Gee Kim ◽  
Junghyun Son ◽  
Changmin Sung
Keyword(s):  
High Throughput ◽  
Doping Analysis ◽  
Gene Doping ◽  
Human Erythropoietin

scholarly journals Legal Basis for the Creation of a System for Preventing Gene Doping and Countering Genetic Modification in Sport

Lex Russica ◽  
2019 ◽  
Vol 1 (9) ◽  
pp. 119-129
Author(s):  
O. A. Shevchenko ◽  
D. I. Vorontsov
Keyword(s):  
Gene Therapy ◽  
Genetic Research ◽  
Significant Risk ◽  
Key Factors ◽  
Potential Harm ◽  
Gene Doping ◽  
Legal Norms ◽  
Genetic Modifications ◽  
Genetic Technologies ◽  
Definition Of

The paper is devoted to the study of the key factors underlying the building of a balanced system of legal norms aimed at preventing gene doping and countering the spread of genetic modifications of athletes. The paper explores the goals of countering doping as such and focuses on the dangers and potential harm to the sports of such a relatively new threat as gene doping. The authors discuss approaches to understanding gene therapy, as well as the prospects of embedding resistance to gene doping in existing legal mechanisms, including the possibility of using the mechanism of therapeutic exceptions (TUE). The current approaches to the definition of the essence of doping are studied and it is determined why gene therapy has a significant risk of being recognized as such. The risks and threats to the sphere of sport caused by the ongoing development of genetic technologies and the spread of their application are outlined. The possible consequences of the use of the results of genetic research, as well as the use of gene therapy, which may affect the field of sports, are discussed. The authors study issues of responsibility for the use of gene doping, subjects against which appropriate sanctions can be imposed, as well as the issue of the application of state coercion to individual subjects, without which the use of gene doping would be impossible. At the same time, the paper formulates the problems that need to be solved in the short term to ensure the preventive nature of the fight against gene doping in sport, and identifies the questions that society must answer to form a system of countering gene doping.

Gene doping

2008 ◽  
Vol 44 ◽  
pp. 125-138 ◽  
Author(s):  
Stephen D.R. Harridge ◽  
Cristiana P. Velloso
Keyword(s):  
Gene Therapy ◽  
Gene Transfer ◽  
Athletic Performance ◽  
Potential Threat ◽  
Altered Expression ◽  
Gene Doping ◽  
Naturally Occurring ◽  
The World ◽  
World Anti Doping Agency ◽  
New Challenges

Gene doping is the misuse of gene therapy to enhance athletic performance. It has recently been recognised as a potential threat and subsequently been prohibited by the World Anti-Doping Agency. Despite concerns with safety and efficacy of gene therapy, the technology is progressing steadily. Many of the genes/proteins which are involved in determining key components of athletic performance have been identified. Naturally occurring mutations in humans as well as gene-transfer experiments in adult animals have shown that altered expression of these genes does indeed affect physical performance. For athletes, however, the gains in performance must be weighed against the health risks associated with the gene-transfer process, whereas the detection of such practices will provide new challenges for the anti-doping authorities.

A duplex qPCR assay for human erythropoietin (EPO) transgene to control gene doping in horses

2020 ◽  
Author(s):  
Hiu Wing Cheung ◽  
Kin‐Sing Wong ◽  
Venus Y.C. Lin ◽  
Terence S.M. Wan ◽  
Emmie N.M. Ho
Keyword(s):  
Qpcr Assay ◽  
Control Gene ◽  
Gene Doping ◽  
Human Erythropoietin
Sign in / Sign up

Export Citation Format

Share Document