Detection of non-targeted transgenes by whole-genome resequencing for gene-doping control

• Published in: Gene therapy Vol. 28; no. 3-4; pp. 199 - 205
• Main Authors: Tozaki, Teruaki, Ohnuma, Aoi, Takasu, Masaki, Nakamura, Kotono, Kikuchi, Mio, Ishige, Taichiro, Kakoi, Hironaga, Hirora, Kei-ichi, Tamura, Norihisa, Kusano, Kanichi, Nagata, Shun-ichi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.04.2021; Nature Publishing Group
• Subjects: 38/23; 631/1647/1513; 631/61/201; Algorithms; Animals; Animals, Genetically Modified; Biomedical and Life Sciences; Biomedicine; Brief Communication; Cell Biology; Cell lines; Doping in horse-racing; Doping in Sports; Erythropoietin; Erythropoietin - genetics; Fibroblasts; Gene Expression; Gene Therapy; Genetic aspects; Genetic engineering; Genetic research; Genetic screening; Genetically modified animals; Genomes; Horses; Human Genetics; Identification and classification; Methods; Nanotechnology; Race horses; Single-nucleotide polymorphism; Transgenes; Japan
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/s41434-020-00185-y

Gene doping has raised concerns in human and equestrian sports and the horseracing industry. There are two possible types of gene doping in the sports and racing industry: (1) administration of a gene-doping substance to postnatal animals and (2) generation of genetically engineered animals by modifying eggs. In this study, we aimed to identify genetically engineered animals by whole-genome resequencing (WGR) for gene-doping control. Transgenic cell lines, in which the erythropoietin gene ( EPO ) cDNA form was inserted into the genome of horse fibroblasts, were constructed as a model of genetically modified horse. Genome-wide screening of non-targeted transgenes was performed to find structural variation using DELLY based on split-read and paired-end algorithms and Control-FREEC based on read-depth algorithm. We detected the EPO transgene as an intron deletion in the WGR data by the split-read algorithm of DELLY. In addition, single-nucleotide polymorphisms and insertions/deletions artificially introduced in the EPO transgene were identified by WGR. Therefore, genome-wide screening using WGR can contribute to gene-doping control even if the targets are unknown. This is the first study to detect transgenes as intron deletions for gene-doping detection.