Toray Industries, Inc.

Tokyo Metropolitan Institute of Medical Science

Tokyo Metropolitan Organization for Medical Research

Other

Japan As Represented by Director-general of National Institute of Infectious Diseases

Ralf Bartenschlager

Bartenschlager, Ralf

Takanobu Kato

Takaji Wakita

Tomoko Date

Michiko Miyamoto

Saburo Sone

Jun-ichi Tanabe

Jun-Ichi Tanabe

Asako Murayama

Jun-Ichi c/o Basic Research Laboratories Tanabe

Tomoko; Date

Michiko; Miyamoto

Saburo; Sone

A nucleic acid includes the 5' untranslated region, a virus protein-coding region including the NS3 protein coding sequence, the NS4A protein coding sequence, the NS4B protein coding sequence, the NS5A protein coding sequence, and the NS5B protein coding sequence, and the 3&#x2032; untranslated region of the HCV J6CF genome in that order from the 5&#x2032; to 3&#x2032; direction. The NS4A protein coding sequence has a mutation for substituting alanine at position 1680 with glutamic acid, as determined on the basis of the amino acid sequence of the precursor protein of the J6CF strain.

Mutant replicon derived from genome of hepatitis C virus J6CF strain

The present invention provides modified hepatitis C virus genomic RNA, comprising nucleotide sequences of genomic RNA portions of two or more types of hepatitis C viruses, which comprises a 5' untranslated region, a core protein coding sequence, an E1 protein coding sequence, a p7 protein coding sequence, an E2 protein coding sequence, an NS2 protein coding sequence, an NS3 protein coding sequence, an NS4A protein coding sequence, an NS4B protein coding sequence, an NS5A protein coding sequence, an NS5B protein coding sequence, and a 3&#x2032; untranslated region, and which can be autonomously replicated. In particular, the present invention relates to modified hepatitis C virus genomic RNA, which can be autonomously replicated by substitution of the RNA sequence portion encoding NS3, NS4, NS5A, and NS5B proteins of hepatitis C virus genomic RNA with a partial RNA sequence encoding NS3, NS4, NS5A, and NS5B proteins of a JFH1 strain shown in SEQ ID NO: 1.

Modified human hepatitis C virus genomic RNA that can be autonomously replicated

The present invention relates to a replicon RNA comprising a nucleotide sequence at least containing the 5' untranslated region, the nucleotide sequence encoding NS3 protein, NS4A protein, NS4B protein, NS5A protein and NS5B protein, and the 3&#x2032; untranslated region on the genomic RNA of hepatitis C virus of genotype 2a.

Nucleic acid construct containing a nucleic acid derived from the genome of hepatitis C virus (HCV) of genotype 2a, and a cell having such nucleic acid construct introduced therein

The present invention provides a method for replicating efficiently an RNA containing fulllength HCV genomic sequence and a method for producing HCV virus particles containing fulllength HCV replicon RNA or fulllength HCV genomic RNA by using a cell culture system. Further, the present invention relates to a method for producing hepatitis C virus particles which comprises culturing a cell, into which a replicon RNA comprising a nucleotide sequence comprising a fulllength genomic RNA sequence of hepatitis C virus of the genotype 2a, at least one selectable marker gene and/or at least one reporter gene and at least one IRES sequence or the fulllength genomic RNA of hepatitis C virus of the genotype 2a is introduced, and generating virus particles in the culture medium. Still further the present invention relates also to a hepatitis C vaccine and an antibody against hepatitis C virus particles.

Nucleic acid construct containing full length genome of human hepatitis C virus, recombinant full length virus genome-replicating cells having the nucleic acid construct transferred thereinto and method of producing hepatitis C virus particle

The present invention relates to a gene derived from a novel fulminant hepatitis C virus strain, an HCV replicon RNA with a high replication efficiency obtained using the gene, and an HCV replicon-replicating cell transfected with the replicon RNA. When the HCV replicon RNA and the HCV replicon-replicating cell of the present invention are used, HCV proteins can be continuously produced in a large amount.

Nucleic acid and gene derived from novel HCV strain and replicon-replicating cell using said gene

Nucleic acid construct containing a nucleic acid derived from the genome of hepatitis C virus (HCV) of genotype 2A, and a cell having such nucleic acid construct introduced therein

The present invention provides a method for replicating efficiently an RNA containing fulllength HCV genomic sequence and a method for producing HCV virus particles containing fulllength HCV replicon RNA or fulllength HCV genomic RNA by using a cell culture system. Further, the present invention relates to a method for producing hepatitis C virus particles which comprises culturing a cell, into which a replicon RNA comprising a nucleotide sequence comprising a fulllength genomic RNA sequence of hepatitis C virus of the genotype 2, at least one selectable marker gene and/or at least one reporter gene and at least one IRES sequence or the fulllength genomic RNA of hepatitis C virus of the genotype 2is introduced, and generating virus particles in the culture medium. Still further the present invention relates also to a hepatitis C vaccine and an antibody against hepatitis C virus particles.

Nucleic acid construct containing fulllength genome of human hepatitis C virus, recombinant fulllength virus genome-replicating cells having the nucleic acid construct transferred thereinto and method of producing hepatitis C virus Particle

Growing community of inventors

Aichi, Japan

Takanobu Kato