Toray Industries, Inc.

Tokyo Metropolitan Institute of Medical Science

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

Tokyo Metropolitan Organization for Medical Research

Jun-ichi Tanabe

Takaji Wakita

Saburo Sone

Tomoko Date

Takanobu Kato

Michiko Miyamoto

Tetsuro Suzuki

Koji Ishii

Tatsuo Miyamura

Ralf Bartenschlager

Ryosuke Suzuki

Takaji c/o Department Of Virology Ii Wakita

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 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 method for producing a recombinant hepatitis C virus-like particle comprising the steps of introducing into (i) a cell in which an RNA replicon comprising a nucleotide sequence comprising the 5' untranslated region, the nucleotide sequence coding for the NS3, NS4A, NS4B, NS5A, and NS5B proteins, and the 3&#x2032; untranslated region of a genome RNA derived from a hepatitis C virus strain autonomously replicates, (ii) a vector expressing the Core, E1, E2, and p7 proteins derived from a hepatitis C virus strain that is the same as or different from that as defined in the above (i), culturing the cell, and recovering the produced virus-like particle, and a recombinant hepatitis C virus particle produced by this method.

Recombinant human hepatitis C virus-like particle and method for producing the same

The present invention relates to a method for producing infectious hepatitis C virus (HCV) particles, comprising a step of introducing an expression vector into a cell that allows HCV proliferation, such expression vector comprising: DNA sequences encoding the 5' untranslated region, structural proteins, and, if necessary, non-structural proteins of HCV and DNA sequences encoding non-structural proteins and the 3&#x2032; untranslated region derived from the HCV JFH1 strain, which are located downstream of a polymerase I promoter; and a DNA fragment containing an RNA polymerase I terminator, which is located further downstream thereof.

High production system for infectious hepatitis C virus particle

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

Kanagawa, Japan

Jun-ichi Tanabe