An Improved Protocol for the Production of Lentiviral Vectors

Logan Y Brown; Wendy Dong; Boris Kantor

doi:10.1016/j.xpro.2020.100152

An Improved Protocol for the Production of Lentiviral Vectors

STAR Protoc. 2020 Oct 27;1(3):100152. doi: 10.1016/j.xpro.2020.100152. eCollection 2020 Dec 18.

Authors

Logan Y Brown¹, Wendy Dong¹, Boris Kantor^{1

2}

Affiliations

¹ Duke Viral Vector Core, Department of Neurobiology, Duke University Medical Center, Durham, NC 27710, USA.
² CLAIRIgene Inc., Durham, NC 27701, USA.

Abstract

Lentiviral vectors are an ideal gene-delivery system for large gene-editing tools, such as the clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 system, due to their high packaging capacity and broad tropism. Here, we present a calcium phosphate-based protocol for lentiviral production and concentration for in vitro and in vivo use. This revised procedure has been optimized to ensure high viral titers and transduction efficiency and is scalable to meet specific production needs.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Genetic Techniques*
Genetic Vectors / metabolism*
HEK293 Cells
Humans
Lentivirus / genetics*
Plasmids / genetics
Transfection
Ultracentrifugation
Virion / metabolism