Open AccessHuman Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
Author(s) -
Porter Samuel S.,
McBride Alison A.
Publication year - 2020
Publication title -
current protocols in microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.813
H-Index - 35
eISSN - 1934-8533
pISSN - 1934-8525
DOI - 10.1002/cpmc.101
Subject(s) - biology , virology , minicircle , papillomaviridae , foreskin , viral replication , dna , virus , cell culture , genetics , cancer , cervical cancer
This protocol describes the production of human papillomavirus (HPV)–derived quasiviruses. Quasiviruses are infectious particles that are produced in 293TT packaging cells and contain a complete viral genome. We describe methods for infection of primary human keratinocytes with HPV quasiviruses, as well as assays to measure early viral DNA replication and transcription. Published 2020. U.S. Government. Basic Protocol 1 : Transfection, harvest, and isolation of HPV quasiviruses Alternate Protocol 1 : Packaging HPV DNA replicated in 293TT cells Alternate Protocol 2 : Production of higher‐purity quasivirus using the “Ripcord” method Support Protocol 1 : Production of HPV minicircles Support Protocol 2 : Production of recircularized HPV genomes Support Protocol 3 : Screening of fractions for viral proteins Support Protocol 4 : Screening of fractions for viral DNA Support Protocol 5 : Measuring viral titer Support Protocol 6 : Quantitation of quasivirions Basic Protocol 2 : Infection of primary human foreskin keratinocytes with quasivirus Basic Protocol 3 : HPV quasivirus transcription assay Basic Protocol 4 : HPV quasivirus replication assay
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