Development of HBV-and/or HDV-susceptible cells, cell lines and non-human animals

Patent metadata

Field	Value
Publication number	US-10618947-B2
Application number	US-201314442279-A
Country	US
Kind code	B2
Filing date	Nov 12, 2013
Priority date	Nov 12, 2012
Publication date	Apr 14, 2020
Grant date	Apr 14, 2020

Abstract

The present invention relates to a novel Hepatitis B virus (HBV) and/or Hepatitis D virus (HDV) receptor and its use for the development of cells, cell lines and non-human animals that are susceptible to HBV and/or HDV infection and can be used for immunological studies and/or for the screening of drugs, post-entry restriction factors and host dependency factors. It further relates to the use of the receptor for the identification of compounds useful in the treatment of HBV and/or HDV infection.

First claim

The invention claimed is:

1. A method for producing a cell that is susceptible to Hepatitis B virus (HBV) and/or Hepatitis D Virus (HDV) infection, or has an increased susceptibility to HBV and/or HDV infection, or is able to bind HBV and/or HDV, said method comprising the steps of:
A) providing a cell that is non-susceptible to HBV and/or HDV infection or has a low susceptibility to HBV and/or HDV infection or is unable to bind HBV and/or HDV:
B) transducing said cell with a viral vector comprising a nucleic acid sequence encoding:
an amino acid sequence comprising SEQ ID NO: 1, 3, 4, 5, 6, 7, or 8 or an amino acid sequence that is at least 90% identical to SEQ ID NO: 1, 3, 4, 5, 6, 7, or 8, and an amino acid sequence consisting of the general formula Pro-Tyr-X-Gly-Ile, wherein X is selected from Lys, Arg and Val,
C) knocking-down one or more endogenous genes of said cell, wherein one of said endogenous genes is the gene encoding the natural sodium taurocholate cotransporter polypeptide (NTCP/SCL10A1) polypcptide of said cell, and said knocking-down one or more endogenous genes of said cell is achieved by means of an shRNA vector; and
D) culturing the transduced cell in the presence of DMSO when the transduced cell is contacted with HBV and/or HDV.

2. A method for producing a cell that is susceptible to Hepatitis B virus (HBV) and/or Hepatitis D Virus (HDV) infection, or has an increased susceptibility to HBV and/or HDV infection, or is able to bind HBV and/or HDV, said method comprising the steps of:
A) providing a cell that is non-susceptible to HBV and/or HDV infection or has a low susceptibility to HBV and/or HDV infection or is unable to bind HBV and/or HDV;
B) transducing said cell with a viral vector comprising a nucleic acid sequence encoding: an amino acid sequence consisting of SEQ ID NO: 2 or an amino acid sequence that is at least 90% identical to SEQ ID NO: 2, and an amino acid sequence comprising the sequence of SEQ ID NO: 1, 3, 4, 5, 6, 7, or 8 or an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 1, 3, 4, 5, 6, 7, or 8,
C) knocking-down one or more endogenous genes of said cell, wherein one of said endogenous genes is the gene encoding the natural sodium taurocholate cotransporter polypeptide (NTCP/SCL10A1) polypeptide of said cell, and said knocking-down one or more endogenous genes of said cell is achieved by means of an shRNA vector; and
D) culturing the transduced cell in the presence of DMSO when the transduced cell is contacted with HBV and/or HDV.

3. A method for producing a cell that is susceptible to Hepatitis B virus (HBV) and/or Hepatitis D Virus (HDV) infection, or has an increased susceptibility to HBV and/or HDV infection, or is able to bind HBV and/or HDV, said method comprising the steps of:
A) providing a cell that is non-susceptible to HBV and/or HDV infection or has a low susceptibility to HBV and/or HDV infection or is unable to bind HBV and/or HDV;
B) introducing into said cell a nucleic acid sequence encoding:
an amino acid sequence consisting of SEQ ID NO: 2 or an amino acid sequence that is at least 90% identical to SEQ ID NO: 2, and
an amino acid sequence comprising the sequence of SEQ ID NO: 1, 3, 4, 5, 6, 7, or 8 or an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 1, 3, 4, 5, 6, 7, or 8.

4. A method for producing a cell that is susceptible to Hepatitis B virus (HBV) and/or Hepatitis D Virus (HDV) infection, or has an increased susceptibility to HBV and/or HDV infection, or is able to bind HBV and/or HDV, said method comprising the steps of:
A) providing a cell that is non-susceptible to HBV and/or HDV infection or has a low susceptibility to HBV and/or HDV infection or is unable to bind HBV and/or HDV;
B) introducing into said cell a nucleic acid sequence encoding:
an amino acid sequence comprising the sequence of SEQ ID NO: 1, 3, 4, 5, 6, 7, or 8, or an amino acid sequence that is at least 90% identical to SEQ ID NO: 1, 3, 4, 5, 6, 7, or 8, and
an amino acid sequence consisting of the general formula Pro-Tyr-X-Gly-Ile, wherein X is selected from Lys, Arg and Val.

Assignees

• Univ Heidelberg Ruprecht Karls

Inventors

• Urban Stephan
• Ni Yi

Classifications

• A01K67/0278

  Knock-in vertebrates, e.g. humanised vertebrates · CPC title

• G01N2333/02

  Hepadnaviridae, e.g. hepatitis B virus · CPC title

• G01N2500/02

  Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor) · CPC title

• C12N2730/10152

  relating to complementing cells and packaging systems for producing virus or viral particles · CPC title

• C12N2730/10122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title