Molmed Spa

Chiara Bovolenta

Paolo Rizzardi

Anna Stornaiuolo

Fulvio Mavilio

Gian Paolo Rizzardi

The present invention provides new stable packaging cell lines and producer cell lines as well as methods to obtain them, and a new method to produce lentiviral vectors using such cell lines. New methods and packaging cell lines of the invention are generated using a baculo-AAV hybrid system for stable expression of structural and regulatory lentiviral proteins, such system comprising a baculoviral backbone containing an integration cassette flanked by AAV ITR, in combination with a plasmid encoding rep protein. This system allows to obtain a stable integration of the structural and regulatory HIV-1 proteins gag/pol and rev. The system allows to obtain a first intermediate including only the structural and regulatory HIV proteins gag/pol and rev, to be used as starting point to obtain stable packaging cell lines as well as producer cell lines.

Stable production of lentiviral vectors

The present invention provides a new semi-stable packaging cell line and a method to produce lentiviral vectors (LV), using the semi-stable packaging cell line. New methods and packaging cell lines of the invention are generated using a baculo-AAV hybrid system for stable expression of structural and regulatory lentiviral proteins, such system comprising a baculoviral backbone containing an integration cassette flanked by AAV ITRs, in combination with a plasmid encoding rep protein. This system allows to obtain a stable integration of the structural and regulatory HIV-1 proteins gag/pol and rev. The system allows to obtain a cell line including the structural and regulatory HIV proteins gag/pol and rev, to be used for a semi-stable LV production.

Semi-stable production of lentiviral vectors

A polynucleotide comprising a nucleotide sequence encoding Vif, wherein each of the amino acids corresponding to positions 127, 128, 130, 131, 132 and 142 of the amino acid sequence in FIG.A representing the Vif wild-type consensus sequence, are replaced with another amino acid, and wherein the nucleotide sequence does not encode the amino acid sequence shown in FIG., representing the Vif sequence of the F12 non-producer variant of HIV-1.

Growing community of inventors

Milan, Italy

Chiara Bovolenta