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Merck
CN

An Improved Protocol for the Production of Lentiviral Vectors.

STAR protocols (2020-12-31)
Logan Y Brown, Wendy Dong, Boris Kantor
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
BES, ≥99.0% (titration)
Sigma-Aldrich
Calcium chloride dihydrate, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Sodium phosphate dibasic, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
Sigma-Aldrich
L-Glutamine, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
Goat Anti-Rabbit IgG Antibody, HRP-conjugate, 1 mg/mL, Upstate®