Abstract
We previously established an infection model for BK polyomavirus (BKPyV) in primary human renal proximal tubule epithelial (RPTE) cells. Use of these cells is limited by their inability to be passaged extensively. Here, we describe RPTE cells immortalized with human telomerase reverse transcriptase (hTERT), which can serve as a model system for acute or persistent BKPyV infection.
Highlights
BK polyomavirus (BKPyV) was first isolated in 1971 (1)
HTERT-expressing lentivirus was produced by cotransfecting the pLenti-CMV-human telomerase reverse transcriptase (hTERT), pRSV-Rev, pMDLg/pRRE, and pMD2.G plasmids into 293TT cells (8, 9)
renal proximal tubule epithelial (RPTE) cells at passage 3 were grown in REGM/REBM medium in a 10-cm dish. hTERT-expressing lentivirus at a multiplicity of infection (MOI) of 0.3 was directly added to the cells and inoculated at 37°C overnight
Summary
BK polyomavirus (BKPyV) was first isolated in 1971 (1). BKPyV is a member of the Polyomaviridae, which is a group of small, icosahedral, nonenveloped viruses with circular double-stranded DNA genomes. HTERT-expressing lentivirus was produced by cotransfecting the pLenti-CMV-hTERT, pRSV-Rev, pMDLg/pRRE, and pMD2.G plasmids into 293TT cells (8, 9). WPRE PGK promoter pLenti-CMV-hTERT-puro CMV promoter hTERT
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