Generation of high-titer pseudotyped lentiviral vectors

Shuang Hu, Mingjie Li, Ramesh Akkina

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

4 Scopus citations

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages125-134
Number of pages10
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1937
ISSN (Print)1064-3745

Keywords

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

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