SLC20A1 knockout to stably express a fusogenic gibbon ape leukemia virus envelope glycoprotein for lentiviral vector production.

ABSTRACT: Lentiviral vectors (LVs) derived from HIV-1 are widely used in gene therapy, with several approved products. However, establishing stable cell lines for constitutive LV production remains a challenge due to the cytotoxicity of vector proteins, including some envelope glycoproteins used for pseudotyping. This study used CRISPR-Cas9 to tackle the fusogenicity-derived cytotoxicity of GaLVΔR and establish a stable cell line for constitutive LV production. By knocking out SLC20A1, the cellular receptor for GaLV, we established 293T SLC20A1 knockout (KO) cells, which support GaLVΔR expression without syncytia formation, while maintaining transient LV titers. Phenotypic characterization of these cells revealed a decrease in cell growth, which could be rescued upon SLC20A2 (a transporter of inorga

SUBMITTER: Nogueira RJ 

PROVIDER: S-EPMC12639841 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature