Virus envelope vector for gene transfer
AssigneeAnGes MG, Inc.
InventorYasufumi Kaneda
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Abstract
A gene transfer vector is prepared by introducing an exogenous gene into an inactivated virus envelope, through a freezing and thawing treatment or mixing with a detergent. There are also provided a pharmaceutical composition for gene therapy containing this gene transfer vector, a kit containing this gene transfer vector, and a gene transfer method employing this gene transfer vector.
Claims31
1-19. (canceled)
A native virus envelope for gene transfer, prepared by a method comprising the steps of:
- (i) purifying the virus;
- and
- (ii) inactivating the virus.
The native virus envelope according to claim 1, wherein the virus envelope is derived from a wild-type virus or a recombinant-type virus.
The native virus envelope according to claim 1, wherein the virus belongs to the Paramyxoviridae family.
The native virus envelope according to claim 1, where in the virus is HVJ.
The native virus envelope according to claim 1, wherein the purification step compromises:
- (a) centrifugation of the virus in a fixed-angle rotor.
The native virus envelop according to claim 1, wherein the purification step comprises:
- (a) centrifugation of the virus in a fixed-angle rotor at about 27,000 g.
A gene transfer vector comprising an exogenous gene encapsulated in a native virus envelope, prepared by a method comprising the steps of:
- mixing the native virus envelope with the exogenous gene in the presence of detergent;
- centrifugation of the mixture;
- and
- recovering a pellet containing the gene transfer vector.
The gene transfer vector according to claim 7, wherein the native virus envelope is prepared by a method comprising the steps of:
- (i) purifying the virus;
- and
- (ii) inactivating the virus.
The gene transfer vector according to claim 8, wherein the purification step comprises:
- (a) centrifugation of the virus in a fixed-angle rotor.
The gene transfer vector according to claim 8, wherein the purification step comprises:
- (a) centrifugation of the virus in a fixed-angle rotor at about 27,000 g.
The gene transfer vector according to claim 7, wherein the detergent is selected from the group consisting of octylglucoside, Triton-X100, CHAPS and NP-40.
The gene transfer vector according to claim 11, wherein the detergent is octylglucoside.
The gene transfer vector according to claim 7, wherein the method further comprises a step of adding protamine sulfate to the exogenous gene.
The use of the gene transfer vector according to claim 7, to introduce an exogenous gene into animal tissue in vivo.
The gene transfer vector according to claim 14, wherein the tissue is selected from the group consisting of liver, skeletal muscle, uterus, brain, eyes, carotid arteries, skin, blood vessels, lung, heart, kidneys, spleen, cancer tissue, nerves, B lymphocytes, and respiratory tract tissue.
A pharmaceutical composition for gene therapy which comprises the gene transfer vector according to claim 7.
A kit for screening gene libraries, which comprises the gene transfer vector according to claim 7.
A method for preparing a native virus envelope for gene transfer, comprising the steps of:
- (i) purifying the virus;
- and
- (ii) inactivating the virus.
The method according to claim 18, wherein the purification step comprises:
- (a) centrifugation of the virus in a fixed-angle rotor.
The method according to claim 18, wherein the purification step comprises:
- (a) centrifugation of the virus in a fixed-angle rotor at about 27,000 g.
A method for preparing a gene transfer vector comprising an exogenous gene encapsulated in a native virus envelope, comprising the steps of:
- mixing the native virus envelope with the exogenous gene in the presence of detergent;
- centrifugation of the mixture;
- and
- recovering a pellet containing the gene transfer vector.
A method for introducing a gene into isolated animal tissue, wherein the method comprises the step of:
- introducing a gene into the isolated animal tissue via the gene transfer vector according to claim 7.
A method for introducing an exogenous gene into a suspended cell, wherein the method comprises the steps of:
- mixing the suspended cell with the gene transfer vector according to claim 7 in the presence of protamine sulfate;
- and
- centrifuging the mixture.
Dependencies are inferred from the claim text.