Retroviral vector hybrids and the use thereof for gene transfer
AssigneeBoehringer Mannheim GmbH
InventorsChristopher Baum, Carol Stocking-Harbers, Wolfram Ostertag
Life of this patent
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Abstract
Replication-defective retroviral vector hybrids which are characterized in that they contain a) the U5 region and/or tRNA primer binding site of MESV as the U5 region and/or tRNA primer binding site in the leader region and b) the U3 and R regions from a Friend murine leukaemia virus (F-MuLV) as the U3 and R regions in the 3'-LTR and which are particularly suitable for an effective gene transfer in haematopoietic stem cells.
Claims26
A retroviral vector hybrid comprising
- a) a 5'-LTR-comprising at least one U5 region or tRNA primer binding site wherein the region or site is selected from the group consisting of the U5 region of MESV, the tRNA primer binding site of MESV, the U5 region of MoMuSV, and the tRNA primer binding site of MoMuSV, and
- b) a 3'-LTR comprising the U3 and R regions from a Friend murine leukaemia virus (F-MuLV).
A vector hybrid selected from the group consisting of pSF1, pSF2, pSF3, and pHM1.
A process for the production of a retrovirally transduced eukaryotic cell which expresses an exogenous gene, said process comprising transducing a eukaryotic cell with a retroviral vector virus, which virus comprises
- a) at least one U5 region or tRNA primer binding site wherein the region or site is selected from the group consisting of the U5 region of MESV, the tRNA primer binding site of MESV, the U5 region of MoMuSV, and the tRNA primer binding site of MoMuSV, and
- b) a 3'-LTR comprising the U3 and R regions from a Friend murine leukaemia virus (F-MuLV).
An eukaryotic cell obtained by the process of transducing a eukaryotic cell with a retroviral vector virus comprising
- a) at least one U5 region or tRNA primer binding site wherein the region or site is selected from the group consisting of the U5 region of MESV, the tRNA primer binding site of MESV, the U5 region of MoMuSV, and the tRNA primer binding site of MoMuSV, and
- b) a 3'-LTR comprising the U3 and R regions from a Friend murine leukaemia virus (F-MuLV).
A replication defective infectious virus particle comprising a retroviral RNA genome, wherein the genome comprises
- a) at least one U5 region or tRNA primer binding site selected from the group consisting of the U5 region of MESV, the tRNA primer binding site of MESV, the U5 region of MoMuSV, and the tRNA primer binding site of MoMuSV, and
- b) further comprises a packaging function, a gene which is heterologous to the virus and can be expressed in the eukaryotic cell, and U3 and R from a Friend murine leukaemia virus (F-MuLV), but
- c) does not comprise active gag, env and pol sequences at the 3' end.
A process for the production of a replication defective infectious virus particle comprising
- a) transfecting a eukaryotic helper cell which has the helper functions gag, env and pol with a vector hybrid comprising
- i) at least one U5 region or tRNA primer binding site wherein the region or site is selected from the group consisting of the U5 region of MESV, the tRNA primer binding site of MESV, the U5 region of MoMuSV, and the tRNA primer binding site of MoMuSV, and
- ii) a 3'-LTR comprising the U3 and R regions from a Friend murine leukaemia virus (F-MuLV),
- b) producing the RNA corresponding to the DNA of the vector hybrid as the virus genome in the cell,
- c) packaging the virus genome into the replication deficient empty virus envelopes formed in the cell, and
- d) isolating the infectious virus particles which contain said virus genome.
A retroviral vector hybrid comprising
- a) the U5 region and tRNA primer binding site of MESV as the U5 region and the tRNA primer binding site in the leader region, and
- b) the U3 and R regions from myeloproliferative sarcoma virus (MPSV) as the U3 and R regions in the 3'-LTR.
A process for the production of a retrovirally transduced eukaryotic cell comprising an exogenous gene, wherein the eukaryotic cell is transduced with the retroviral vector virus comprising:
- a) the U5 region and/or tRNA primer binding site of MESV as the U5 region and/or tRNA primer binding site in the leader region,
- b) the U3 and R regions from myeloproliferative sarcoma virus (MPSV) as the U3 and R regions in the 3'-LTR and
- c) the said exogenous gene.
A replication defective infectious virus particle comprising retroviral RNA as the genome, wherein the genome comprises the U5 regions and/or tRNA primer binding site of MESV as the U5 region and/or tRNA primer binding site, a packaging function and a gene which is heterologous for the virus and can be expressed in the eukaryotic cell, and U3 and R from the myeloproliferative sarcoma virus (MPSV) but no active gag, env and pol sequences at the 3' end.
Dependencies are inferred from the claim text.