Packaging cell lines for pseudotyped retroviral vectors

US5739018A · kind A · utility

Assignees

• The Regents of the University of California · US
• City of Hope · US

Inventors

• Atsushi Miyanohara · San Diego, US
• Jiing-Kuan Yee · Arcadia, US
• Shin-Tai Chen · San Diego, US
• Charles E. Prussak · San Diego, US
• Theodore Friedmann · Village of La Jolla, US

Key dates

• Filing date: Aug 7, 1996
• Grant date: Apr 14, 1998
• Expiry date: Aug 7, 2016

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2840/203
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention features packaging cell lines and recombinant retroviral particles produced thereby, particularly pseudotyped retroviral particles. Preferably, the packaging cell lines are derived from HeLa, Cf2Th, D17, MDCK, or BHK cells, most preferably from Cf2Th cells. Retroviral particles are produced by inducibly expressing an envelope protein of interest (e.g., a retroviral envelope or the envelope protein of vesicular stomatitis virus (VSV G)). Inducible expression of the envelope protein is accomplished by operably linking an envelope protein-encoding nucleotide sequence to an inducible promoter (e.g., a promoter composed of a minimal promoter linked to multiple copies of tetO, the binding site for the tetracycline repressor (tetR) of the Escherichia coli, tetracycline resistance operon Tn10). Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription from the inducible promoter (e.g., a prokaryotic tetracycline repressor polypeptide (tetR)), a transcriptional activation domain, and a second ligand-binding domain (e.g., a ligand-binding domain of a s…