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Viruses2023; 15(9); 1942; doi: 10.3390/v15091942

CRISPR/Cas9-Mediated Targeting of BPV-1-Transformed Primary Equine Sarcoid Fibroblasts.

Abstract: Equine sarcoids (EqS) are fibroblast-derived skin tumors associated with bovine papillomavirus 1 and 2 (BPV-1 and -2). Based on Southern blotting, the BPV-1 genome was not found to be integrated in the host cell genome, suggesting that EqS pathogenesis does not result from insertional mutagenesis. Hence, CRISPR/Cas9 implies an interesting tool for selectively targeting BPV-1 episomes or genetically anchored suspected host factors. To address this in a proof-of-concept study, we confirmed the exclusive episomal persistence of BPV-1 in EqS using targeted locus amplification (TLA). To investigate the CRISPR/Cas9-mediated editing of BPV-1 episomes, primary equine fibroblast cultures were established and characterized. In the EqS fibroblast cultures, CRISPR-mediated targeting of the episomal E5 and E6 oncogenes as well as the BPV-1 long control region was successful and resulted in a pronounced reduction of the BPV-1 load. Moreover, the deletion of the equine Vimentin , which is highly expressed in EqS, considerably decreased the number of BPV-1 episomes. Our results suggest CRISPR/Cas9-based gene targeting may serve as a tool to help further unravel the biology of EqS pathogenesis.
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Publication Date: 2023-09-17 PubMed ID: 37766348PubMed Central: PMC10536948DOI: 10.3390/v15091942Google Scholar: Lookup
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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.

The research explored the use of CRISPR/Cas9 gene editing tool in selectively targeting bovine papillomavirus 1 (BPV-1) – a cause for skin tumors in horses. The study found a significant reduction in BPV-1 load using this method, and also confirmed that BPV-1 doesn’t integrate with the host genome.

Overview of the Research

  • Equine sarcoids (EqS) are skin tumors derived from fibroblasts and are usually linked with bovine papillomavirus types 1 and 2 (BPV-1 and -2). The researchers discovered that the BPV-1 genome wasn’t integrated with the host cell’s genome.
  • Due to this, they proposed the use of CRISPR/Cas9, a gene editing tool, to specifically target the BPV-1 episomes and possibly the suspected host factors that were anchored genetically.

CRISPR/Cas9 Techniques Used in the Study

  • The team used a technique called targeted locus amplification (TLA) to confirm the episomal persistence of BPV-1 in equine sarcoids.
  • We proceeded to evaluate the CRISPR/Cas9-mediated editing of the BPV-1 episomes, establishing and characterizing primary equine fibroblast cultures in the process.

Results of the Study

  • Among the EqS fibroblast cultures, the team performed successful CRISPR-mediated targeting of episomal E5 and E6 oncogenes, as well as the BPV-1 long control region, leading to a significant decrease of the BPV-1 load.
  • The experiment also showed that the removal of equine vimentin, a protein that is overexpressed in eqS, led to a substantial decrease in the number of BPV-1 episomes.
  • The results indicate that CRISPR/Cas9-based gene targeting could be a useful tool to unravel the biology of EqS pathogenesis.

Cite This Article

APA
Monod A, Koch C, Jindra C, Haspeslagh M, Howald D, Wenker C, Gerber V, Rottenberg S, Hahn K. (2023). CRISPR/Cas9-Mediated Targeting of BPV-1-Transformed Primary Equine Sarcoid Fibroblasts. Viruses, 15(9), 1942. https://doi.org/10.3390/v15091942

Publication

Viruses
ISSN: 1999-4915
NlmUniqueID: 101509722
Country: Switzerland
Language: English
Volume: 15
Issue: 9
PII: 1942

Researcher Affiliations

Monod, Anne
  • Swiss Institute of Equine Medicine (ISME), Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.
  • Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.
Koch, Christoph
  • Swiss Institute of Equine Medicine (ISME), Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.
Jindra, Christoph
  • Research Group Oncology, University Equine Clinic, University of Veterinary Medicine, 1210 Vienna, Austria.
Haspeslagh, Maarten
  • Department of Large Animal Surgery, Anesthesiology and Orthopaedics, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium.
Howald, Denise
  • Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.
Wenker, Christian
  • Zoo Basel, Binningerstrasse 40, 4054 Basel, Switzerland.
Gerber, Vinzenz
  • Swiss Institute of Equine Medicine (ISME), Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.
Rottenberg, Sven
  • Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.
Hahn, Kerstin
  • Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.

MeSH Terms

  • Animals
  • Horses
  • CRISPR-Cas Systems
  • Skin Neoplasms
  • Oncogenes
  • Fibroblasts
  • Gene Targeting

Conflict of Interest Statement

All authors were applied at academic institutions during the time the study was performed.

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Citations

This article has been cited 1 times.
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