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Measurement of BK-polyomavirus Non-Coding Control Region Driven Transcriptional Activity Via Flow Cytometry
Published on: July 13, 2019
Efficient propagation of archetype BK and JC polyomaviruses
Nicole M Broekema1, Michael J Imperiale
1Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109-5620, USA. broekenm@umich.edu
Virology
|November 22, 2011
Summary
Archetype BKPyV and JCPyV viruses can now be studied in cell culture. Overexpressing large T antigen in 293TT cells enables the replication of these ubiquitous human pathogens.
Area of Science:
- Virology
- Molecular Biology
- Immunocompromised Host Research
Background:
- BKPyV and JCPyV are ubiquitous human polyomaviruses causing disease in immunocompromised individuals.
- Distinct regulatory regions define archetype and rearranged forms of these viruses.
- Current cell culture systems are inadequate for studying archetype polyomaviruses.
Purpose of the Study:
- To develop a robust cell culture system for propagating archetype BKPyV and JCPyV.
- To investigate the role of large T antigen (TAg) in archetype polyomavirus replication.
Main Methods:
- Utilized 293TT cells, which are human embryonic kidney cells engineered to overexpress SV40 large T antigen.
- Inoculated 293TT cells with archetype BKPyV and JCPyV.
- Monitored viral replication and analyzed the structural integrity of the viral regulatory regions.
Main Results:
- Achieved efficient propagation of archetype BKPyV and JCPyV in 293TT cells.
- Confirmed that the archetypal regulatory region structure was maintained during replication.
- Observed no significant replication for Merkel cell, KI, or WU polyomaviruses under the same conditions.
Conclusions:
- Overexpression of large T antigen in 293TT cells successfully supports the replication of archetype BKPyV and JCPyV.
- This novel cell culture system facilitates detailed study of archetype polyomaviruses.
- The findings pave the way for further research into the pathogenesis and treatment of diseases caused by these viruses.
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