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Drug testing and analysis2022; 14(8); 1429-1437; doi: 10.1002/dta.3267

Use of mitochondrial sequencing to detect gene doping in horses via gene editing and somatic cell nuclear transfer.

Abstract: Gene editing and subsequent cloning techniques offer great potential not only in genetic disease correction in domestic animals but also in livestock production by enhancement of desirable traits. The existence of the technology, however, leaves it open to potential misuse in performance-led sports such as horseracing and other equestrian events. Recent advances in equine gene editing, regarding the generation of gene-edited embryos using CRISPR/Cas9 technology and somatic cell nuclear transfer, have highlighted the need to develop tools to detect potential prohibited use of the technology. One possible method involves the characterisation of the mitochondrial genome (which is not routinely preserved during cloning) and comparing it with the sequence of the registered dam. We present here our approach to whole-mitochondrial sequencing using tiled long-range PCR and next-generation sequencing. To determine whether the background mutation rate in the mitochondrial genome could potentially confound results, we sequenced 10 sets of dam and foal duos. We found variation between duos but none within duos, indicating that this method is feasible for future screening systems. Analysis of WGS data from over 100 Thoroughbred horses revealed wide variation in the mitochondria sequence within the breed, further displaying the utility of this approach.
© 2022 John Wiley & Sons, Ltd.
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Publication Date: 2022-04-25 PubMed ID: 35362263DOI: 10.1002/dta.3267Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates the use of mitochondrial sequencing to detect illegal gene doping in horses through gene editing and cloning. The researchers conducted whole-mitochondrial sequencing on horse samples to identify any difference in their genetic structures as a result of prohibited gene editing practices.

Background of the Research

  • The application of gene-editing and cloning techniques in domestic animals and livestock offers potential benefits in rectifying genetic diseases and enhancing desired traits.
  • However, the presence of such technology can lead to misuse in performance-oriented sports like horse racing and equestrian events where individuals may seek unfair competitive advantages.
  • Gene-edited embryos created from the CRISPR/Cas9 technology and somatic cell nuclear transfer in horses have intensified the call for the development of detection tools against the unauthorized use of such technology.

Research Methodology

  • The research proposed the use of a method involving the organization of the mitochondrial genome, which typically isn’t preserved during cloning, and comparing it with the sequence of the registered female parent.
  • Utilizing tiled long-range PCR and next-generation sequencing, the scientists sequenced the whole mitochondrial genome.
  • To ascertain whether or not the background mutation rate in the mitochondrial genome can potentially influence results, the researchers sequenced 10 pairings of dams and their young.

Research Findings

  • The research revealed variations among the dam-foal duos but none within the duos, demonstrating that the mitochondrial sequencing method could be viable for future screening systems.
  • Furthermore, by analyzing Whole Genome Sequencing (WGS) data from over 100 Thoroughbred horses, there was a wide variation within the same breed in the mitochondrial sequences, demonstrating that the method described has a practical use for differentiating individual horses.

Implication of the Research

  • This research offers a promising approach to detecting potential gene doping in the sport of horse racing and equestrian events.
  • By distinguishing individual differences at the genetic level through mitochondrial sequencing, the detection of gene doping becomes feasible, encouraging fairness and integrity in equestrian competitions.

Cite This Article

APA
Maniego J, Pesko B, Habershon-Butcher J, Hincks P, Taylor P, Tozaki T, Ohnuma A, Stewart G, Proudman C, Ryder E. (2022). Use of mitochondrial sequencing to detect gene doping in horses via gene editing and somatic cell nuclear transfer. Drug Test Anal, 14(8), 1429-1437. https://doi.org/10.1002/dta.3267

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English
Volume: 14
Issue: 8
Pages: 1429-1437

Researcher Affiliations

Maniego, Jillian
  • Sport and Specialised Analytical Services, LGC, Fordham, UK.
Pesko, Bogusia
  • Sport and Specialised Analytical Services, LGC, Fordham, UK.
Habershon-Butcher, Jocelyn
  • British Horseracing Authority, London, UK.
Hincks, Pamela
  • Sport and Specialised Analytical Services, LGC, Fordham, UK.
Taylor, Polly
  • Sport and Specialised Analytical Services, LGC, Fordham, UK.
Tozaki, Teruaki
  • Genetic Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Japan.
Ohnuma, Aoi
  • Genetic Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Japan.
Stewart, Graham
  • School of Biosciences and Medicine, University of Surrey, Guildford, UK.
Proudman, Christopher
  • School of Veterinary Medicine, University of Surrey, Guildford, UK.
Ryder, Edward
  • Sport and Specialised Analytical Services, LGC, Fordham, UK.

MeSH Terms

  • Animals
  • CRISPR-Cas Systems
  • Doping in Sports
  • Gene Editing / methods
  • Gene Editing / veterinary
  • Horses / genetics
  • Mitochondria / genetics
  • Nuclear Transfer Techniques / veterinary

Grant Funding

  • British Horseracing Authority

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Citations

This article has been cited 2 times.
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