Optimization of human papillomavirus-based pseudovirus techniques for efficient gene transfer

Timra D Gilson,Elliot J Androphy,Ryan T Gibson

doi:10.1038/s41598-020-72027-1

Timra D Gilson, Elliot J Androphy + Show 1 more

Open Access

https://doi.org/10.1038/s41598-020-72027-1

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Abstract

Human papillomavirus (HPV) L1 and L2 capsid proteins self-assemble into virions capable of efficiently packaging either its 8 kilobase genome or non-viral DNA. The ability of HPV capsids to package non-viral DNA makes these a useful tool for delivering plasmids to study proteins of interest in a variety of cell types. We describe optimization of current methods and present new protocols for using HPV capsids to deliver non-viral DNA thereby providing an alternative to DNA transfection. Using keratinocyte generated extracellular matrices can enhance infection efficiency in keratinocytes, hepatocytes and neuronal cells. Furthermore, we describe a suspension-based efficient technique for infecting different cell types.

Highlights

Human papillomavirus (HPV) L1 and L2 capsid proteins self-assemble into virions capable of efficiently packaging either its 8 kilobase genome or non-viral DNA
We examine and compare current methods used in HPV QsV/PsV production and infection, as well as identify improved conditions to mediate delivery of plasmids via PsV into multiple cell types
extracellular matrix (ECM) derived from keratinocyte cell lines NIKS, N/TERT, and HaCaT cells promoted high infectivity compared to no ECM (Fig. 4)

Summary

Introduction

Human papillomavirus (HPV) L1 and L2 capsid proteins self-assemble into virions capable of efficiently packaging either its 8 kilobase genome or non-viral DNA. We describe a new infection method for HPV PsV that efficiently mediates gene transfer in difficult to transfect cell lines. PsV infections of N/TERT and HaCaT cells were dramatically reduced compared to HEK293 TT and HeLa cells (Fig. 1B). Both NH4OH and EDTA methods generated ECM able to bind PsV and promote infection of HaCaT cells (Fig. 1D).

Results

Conclusion