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Production of Lentiviral Vectors

a technology of lentiviral vectors and lentiviral peptides, which is applied in the field of production of lentiviral vectors, can solve the problems of constitutive vector production, difficult production of replication-defective lentiviral vectors for large-scale clinical use, and time-consuming transient transfection systems for virus production

Inactive Publication Date: 2010-01-07
TRIZELL LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production of replication-defective lentiviral vectors for large-scale clinical use is challenging.
As transient transfection systems for virus production can be problematic and time-consuming, attempts have been made to develop stable large-scale production systems.
However, the toxicity of lentiviral protease and the fusogenic envelope protein, VSV-G, has prohibited constitutive vector production.
In addition, baculoviruses are easy to produce in large-scale and high titers, and they present few safety problems as they are not able to replicate in mammalian cells.

Method used

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  • Production of Lentiviral Vectors
  • Production of Lentiviral Vectors

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Embodiment Construction

[0011]To perform the method of the invention, the baculovirus(es) must comprise a lentivirus transfer construct, gag, pol, a suitable envelope protein and rev.

[0012]The term “lentivirus transfer construct” will be known to those skilled in the art. A lentivirus transfer construct is a source of lentiviral RNA genome / lentiviral vector RNA and transgene cassette. One example of a lentivirus transfer construct is LV1-GFP. Methods for producing lentivirus transfer constructs will also be known to those skilled in the art.

[0013]The term “envelope protein” will be known to those skilled in the art. An envelope protein is a protein that protects the nucleic acid of a virus. An example of envelope protein suitable for use, in the invention is VSV-G.

[0014]The term “producer cell” will be known to those skilled in the art. Examples of producer cells suitable for use in the invention are 293T, HepG2, CHO, BHK, Sf9, Sf21, 293, BTI-Tn 5 B 1-4, COS, NIH / 3T3, Vero, NSO or PerC6 cells. The produced...

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Abstract

The present invention is a method of generating a lentivirus vector, comprising cloning each of a leotivimus transfer construct, gag, pol, an envelope protein and rev respectively into the same or different baculoviruses, and transducing a producer cell with the or each baculovirus.

Description

FIELD OF THE INVENTION[0001]The invention relates to methods for the production of lentiviral vectors.BACKGROUND OF THE INVENTION[0002]Lentiviruses, such as Human Immunodeficiency Virus I, are promising tools for gene therapy due to their ability to transduce and integrate into the genome of both dividing and non-dividing cells. However, the production of replication-defective lentiviral vectors for large-scale clinical use is challenging. Lentiviral vectors are normally produced by cotransfecting 293T human embryonic kidney cells with several different plasmid constructs. The first clinical lentiviral vector production was based on a two-plasmid system (Lu et al., 2004). To further improve the safety of the system, lentivirus genome can be separated into four plasmids. The plasmids are a self-inactivating transfer vector; a packaging plasmid containing gag-pol; a rev plasmid; and an envelope glycoprotein plasmid which usually encodes vesicular stomatitis virus glycoprotein G (VSV-G...

Claims

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Application Information

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IPC IPC(8): C12N15/79
CPCA61K48/0091C07K14/005C12N15/86C12N2810/6081C12N2740/16043C12N2740/16045C12N2740/16051C12N2740/16022C12N15/79C12N15/64A61K48/00C12N2710/14051C12N7/00C12N2710/14021C12N2710/14043C12N2740/15043C12N2740/15051
Inventor LESCH, HANNA P.AIRENNE, KARI J.YLA-HERTTUALA, SEPPO
Owner TRIZELL LTD
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