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Generation of High-Titer Pseudotyped Lentiviral Vectors

  • Shuang Hu
  • Mingjie Li
  • Ramesh AkkinaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1937)

Abstract

Lentiviral vectors (LVs) are widely used in gene transfer protocols due to many advantages that include stable gene expression, higher transgene payloads, and, importantly, the ability to pseudotype the vectors with a diverse number of heterologous viral envelopes with broad or restricted cell tropism depending on the need. The pseudotyping process also allows for incorporation of specific antibodies/ligands to engineer LVs. These features greatly facilitate customization of lentiviral vectors for cell/tissue specific gene delivery. The VSV-G protein containing envelope remains the most widely used among the viral glycoproteins used for LV pseudotyping due to its versatile host range and stability. However, many other viral envelopes are being identified for special applications of LVs. Here we describe the methodology to generate pseudotyped LVs using a four-plasmid transient transfection system focusing on aspects to generate high-titer vector stocks.

Key words

Lentiviral vector Viral vector pseudotyping Vector pseudotyping with VSV-G Vector production and concentration Vector titration 

Notes

Acknowledgments

Work reported here was supported by NIH grants to RA. We would like to thank Lauren Kinner-Bibeau for assistance in manuscript preparation.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Medical Microbiology & ImmunologyUniversity of CaliforniaDavisUSA
  2. 2.Department of Neurology and Hope Center for Neurological DisordersWashington University School of MedicineSt LouisUSA
  3. 3.Department of Microbiology, Immunology and PathologyColorado State UniversityFort CollinsUSA

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