US 5,670,354 A · Use of VSV-G pseudotyped vectors for transfer of genes into embryos

Abstract

The present application discloses retrovirus-derived vectors in which the retroviral envelope glycoprotein has been replaced by the G glycoprotein of vesicular stomatitis virus, and the use of these vectors in the transfer of exogenous genes into the cells of a wide variety of non-mammalian organisms. Also disclosed is a method for the generation of retroviral vectors in high titers, wherein a recombinant, stable host cell line is provided which harbors the retroviral vector of interest without envelope protein. High-titer retroviral vector production is initiated by introducing nucleic acid encoding a functional membrane-associated protein into the cell line. The vectors disclosed in the present application can be concentrated by ultracentrifugation to titers greater than 10.sup.9 cfu/ml which are especially useful in human gene therapy trials, and can also infect cells, such as hamster and fish cells, that are ordinarily resistant to infection with vectors containing the retroviral envelope protein.

Claims19

Dependent claims

A method of introducing heterologous nucleic acid into a primordial germ cell of a non-mammalian animal species selected from the group consisting of an amphibian, an insect and a fish, comprising:

exposing embryos of an individual member of said non-mammalian animal species to a vector particle, said vector particle comprising:
said heterologous nucleic acid;
a nucleocapsid encapsidating said nucleic acid, said nucleocapsid comprising nucleocapsid protein of Moloney murine leukemia virus (MoMLV);
a membrane surrounding said nucleocapsid, said membrane having VSV G protein associated therewith, whereby said vector particles are introduced into the genome of said primordial germ cell of said embryo;
growing said embryos into adults;
breeding said adults to produce an F.sub.1 generation;
and
identifying adults of said F.sub.1 generation that produce an F.sub.2 generation carrying said heterologous nucleic acid, wherein said adults contain said heterologous nucleic acid in their germ line.

5 more elements

Dependencies are inferred from the claim text.

Use of VSV-G pseudotyped vectors for transfer of genes into embryos

Life of this patent

Use of VSV-G pseudotyped vectors for transfer of genes into embryos

Life of this patent

Abstract

Claims19