1. Identification of African Elephant Polyomavirus in wild elephants and the creation of a vector expressing its viral tumor antigens to transform elephant primary cells
- Author
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Orkide O. Koyuncu, Virginia R. Pearson, Joshua D. Schiffman, Glenn F. Rall, Rosann Robinson, Cristhian Toruno, Julian Scherer, Lisa M. Abegglen, Jens B. Bosse, and Lynn W. Enquist
- Subjects
Biopsy ,Elephants ,DNA cloning ,Artificial Gene Amplification and Extension ,Genome ,Polymerase Chain Reaction ,Epithelium ,law.invention ,0403 veterinary science ,law ,Animal Cells ,Medicine and Health Sciences ,Vector (molecular biology) ,Antigens, Viral, Tumor ,Polymerase chain reaction ,Mammals ,0303 health sciences ,Multidisciplinary ,biology ,Captive elephants ,Eukaryota ,04 agricultural and veterinary sciences ,Oncology ,Vertebrates ,Medicine ,Cellular Types ,Anatomy ,Polyomavirus ,Research Article ,040301 veterinary sciences ,Science ,Animals, Wild ,Surgical and Invasive Medical Procedures ,Genome, Viral ,Molecular cloning ,DNA construction ,Transfection ,Research and Analysis Methods ,African elephant ,03 medical and health sciences ,biology.animal ,Animals ,Molecular Biology Techniques ,Gene ,Molecular Biology ,030304 developmental biology ,Polyomavirus Infections ,Organisms ,Endothelial Cells ,Biology and Life Sciences ,Cancers and Neoplasms ,Epithelial Cells ,Cell Biology ,Virology ,Transformation (genetics) ,Tumor Virus Infections ,Biological Tissue ,Amniotes ,Plasmid Construction ,Zoology ,Cloning - Abstract
Wild elephant populations are declining rapidly due to rampant killing for ivory and body parts, range fragmentation, and human-elephant conflict. Wild and captive elephants are further impacted by viruses, including highly pathogenic elephant endotheliotropic herpesviruses. Moreover, while the rich genetic diversity of the ancient elephant lineage is disappearing, elephants, with their low incidence of cancer, have emerged as a surprising resource in human cancer research for understanding the intrinsic cellular response to DNA damage. However, studies on cellular resistance to transformation and herpesvirus reproduction have been severely limited, in part due to the lack of established elephant cell lines to enable in vitro experiments. This report describes creation of a recombinant plasmid, pAelPyV-1-Tag, derived from a wild isolate of African Elephant Polyomavirus (AelPyV-1), that can be used to create immortalized lines of elephant cells. This isolate was extracted from a trunk nodule biopsy isolated from a wild African elephant, Loxodonta africana, in Botswana. The AelPyV-1 genome contains open-reading frames encoding the canonical large (LTag) and small (STag) tumor antigens. We cloned the entire early region spanning the LTag and overlapping STag genes from this isolate into a high-copy vector to construct a recombinant plasmid, pAelPyV-1-Tag, which effectively transformed primary elephant endothelial cells. We expect that the potential of this reagent to transform elephant primary cells will, at a minimum, facilitate study of elephant-specific herpesviruses.
- Published
- 2020