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

PeerJ
Kai AokiKazuhiro Takekoshi

Abstract

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. 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. 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 d...Continue Reading

References

Jul 24, 1998·Clinical Chemistry and Laboratory Medicine : CCLM·C OrlandoM Pazzagli
Jun 9, 2001·The New England Journal of Medicine·D A RothUNKNOWN Factor VIII Transkaryotic Therapy Study Group
Jul 19, 2008·Hepatology Research : the Official Journal of the Japan Society of Hepatology·Hironari KanemuraJiro Fujimoto
Nov 17, 2010·The Journal of Physiology·Wolfgang Jelkmann
Oct 18, 2014·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jerry R MendellBrian K Kaspar
Feb 18, 2016·Medicine and Science in Sports and Exercise·Jeff M Baker, Gianni Parise
Jan 26, 2017·Molecular and Cellular Endocrinology·Olivier SalaminNicolas Leuenberger
Dec 30, 2017·Human Gene Therapy·Shao Wen XiaoShan Wen Zhang
Aug 10, 2018·Molecular Pharmaceutics·Takaaki ItoHirokazu Okamoto
Sep 7, 2018·Medicine and Science in Sports and Exercise·Diresibachew W HaileYannis P Pitsiladis

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Methods Mentioned

BETA
blood collection
PCR
electrophoresis

Software Mentioned

GraphPad Prism
CLC Sequence Viewer
GraphPad

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