A DNA recombination-based approach to eliminate papillomavirus infection.
At present, no treatments exist that effectively target and eliminate papillomaviruses (PVs) from infected cells or prevent its replication. We are employing a strategy to prevent virus replication in PV-infected cells through the conditional expression of the herpes simplex virus type 1 thymidine kinase (TK) gene. Expression of TK in this system is expected to be triggered by a homologous recombination event between the endogenous PV genome and a nonexpressing TK gene cassette. Recombination between these two DNAs is expected to change the nonexpressing cassette into a form that expresses TK. Various constructs were generated to express the TK in the above manner. Transfection of cell lines with a TK nonexpressing plasmid did not result in TK production due to alternative splicing and polyadenylation site selection. However, cotransfection of cell lines with PV plasmids along with the above TK construct containing short segments of PV sequences resulted in a recombination event that led to TK expression as shown by Northern and Western blot analyses. We also developed a TK expression cassette utilizing an adeno-associated virus (AAV) vector. Delivery of the cassette by AAV to PV-infected cells resulted in TK expression, and ganciclovir treatment resulted in efficient killing of these cells.
Carson
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A
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Wang
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Z
Xiao
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Khan
S A
SA
D000998:Antiviral Agents / Q000627:therapeutic use; D015152:Blotting, Northern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage*; D000229:Dependovirus / Q000235:genetics; D005260:Female; D015774:Ganciclovir / Q000627:therapeutic use; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D018259:Herpesvirus 1, Human / Q000201:enzymology*; D006801:Humans; D030361:Papillomavirus Infections / Q000628:therapy* / Q000821:virology; D011995:Recombination, Genetic; D013937:Thymidine Kinase / Q000032:analysis / Q000235:genetics*; D002583:Uterine Cervical Neoplasms / Q000628:therapy* / Q000821:virology; D014779:Virus Replication
D000998:Antiviral Agents; D004247:DNA; D013937:Thymidine Kinase; D015774:Ganciclovir