US5585254A — Autonomous parvovirus gene delivery vehicles and …

Claim 1

Independent Claim 72 words

A recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterolegous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 2

Dependent Claim 72 words

A recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterolegous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 3

Dependent Claim 39 words

The vector of claim 1, wherein said heterologous nucleic acid sequence is selected from the group consisting of a heterologous control element and a heterologous coding region, said heterologous control element being operably linked to said heterologous coding region.

Claim 4

Dependent Claim 15 words

The vector of claim 1, wherein said heterologous control element comprises a heterologous response element.

Claim 5

Dependent Claim 23 words

The vector of claim 1, wherein said heterologous nucleic acid sequence comprises a heterologous control element operatively linked to a heterologous coding region.

Claim 6

Dependent Claim 37 words

The vector of claim 1, wherein said heterologous nucleic acid sequence comprises at least one heterologous response element operatively linked to a promoter selected from the group consisting of an autonomous parvovirus promoter and a heterologous promoter.

Claim 7

Dependent Claim 35 words

The vector of claim 2, wherein said heterologous control element is operatively linked to at least one coding region selected from the group consisting of an autonomous parvovirus coding region and said heterologous coding region.

Claim 8

Dependent Claim 70 words

The vector of claim 2, wherein said heterologous coding region is operatively linked to a transcription control sequence selected from the group consisting of an autonomous parvovirus transcription control sequence that regulates the expression of parvovirus nonstructural polypeptide genes, an autonomous parvovirus transcription control sequence that regulates the expression of parvovirus structural polypeptide genes, and a heterologous transcription control sequence comprising a promoter and at least one heterologous response element.

Claim 9

Dependent Claim 14 words

The vector of claim 1, wherein said vector is packaged into a virus particle.

Claim 10

Dependent Claim 51 words

The vector of claim 1, wherein said parvovirus nucleic acid sequences are selected from the group consisting of LuIII parvovirus, minute virus of mice, hamster parvovirus, feline panleukopenia virus, canine parvovirus, porcine parvovirus, latent rat virus, mink enteritis virus, human parvovirus, bovine parvovirus, and Aleutian mink disease parvovirus nucleic acid sequences.

Claim 11

Dependent Claim 37 words

The vector of claim 1, wherein said parvovirus nucleic acid sequences are selected from the group consisting of LuIII parvovirus, minute virus of mice MVMi, minute virus of mice MVMp, and hamster parvovirus Hi nucleic acid sequences.

Claim 12

Dependent Claim 18 words

The vector of claim 1, wherein said parvovirus nucleic acid sequences comprise a LuIII parvovirus nucleic acid sequence.

Claim 13

Dependent Claim 21 words

The vector of claim 2, wherein said heterologous coding region is operatively linked to an autonomous parvovirus P4 transcription control sequence.

Claim 14

Dependent Claim 20 words

The vector of claim 2, wherein said heterologous coding region is operatively linked to a LuIII P4 transcription control sequence.

Claim 15

Dependent Claim 36 words

The vector of claim 1, wherein said heterologous nucleic acid sequence is selected from the group consisting of a cell-selective response element, a hormone receptor response element, an antibiotic response element, and a carbohydrate response element.

Claim 16

Dependent Claim 32 words

The vector of claim 14, wherein said cell-selective response element is capable of being activated by a trans-activating regulatory element selectively produced in a cell type to which said vector is targeted.

Claim 17

Dependent Claim 27 words

The vector of claim 15, wherein said cell type is selected from the group consisting of a cancer cell and a cell infected by an infectious agent.

Claim 18

Dependent Claim 76 words

The vector of claim 1, wherein said heterologous nucleic acid sequence is selected from the group consisting of a tetracycline response element, a GAL4 response element, a progesterone receptor response element, a glucocorticoid receptor response element, an immunoglobulin kappa light chain enhancer, an immunoglobulin heavy chain enhancer, an α-1-antitrypsin enhancer, a serum albumin enhancer, a chorionic gonadotropin α-chain enhancer, a chorionic gonadotropin β-chain enhancer, an IL-2 enhancer, an IL-2 receptor enhancer, and an HIV response element.

Claim 19

Dependent Claim 33 words

The vector of claim 1, wherein said heterologous nucleic acid sequence encodes a functional protein selected from the group consisting of a cytotoxic agent, an immunopotentiator, a vaccine antigen and functional equivalents thereof.

Claim 20

Dependent Claim 60 words

The vector of claim 1, wherein said heterologous nucleic acid sequence encodes a functional protein selected from the group consisting of a diphtheria toxin, a ricin toxin, a modeccin toxin, an abrin toxin, a Pseudomonas exotoxin, a shiga toxin, a pokeweed antiviral protein, α-amanitin, a ribosome inhibiting protein, an autonomous parvovirus nonstructural protein, HSV thymidine kinase, and functional equivalents thereof.

Claim 21

Dependent Claim 37 words

The vector of claim 1, wherein said heterologous nucleic acid sequence encodes a functional protein selected from the group consisting of a diphtheria A-chain toxin, an autonomous parvovirus NS1 protein, HSV thymidine kinase, and functional equivalents thereof.

Claim 22

Dependent Claim 30 words

The vector of claim 1, wherein said heterologous nucleic acid sequence encodes a functional RNA selected from the group consisting of an antisense RNA, a ribozyme, and an RNA-based drug.

Claim 23

Dependent Claim 15 words

The vector of claim 1, wherein said heterologous nucleic acid sequence encodes a marker protein.

Claim 24

Dependent Claim 60 words

The vector of claim 1, wherein said parvovirus nucleic acid sequences comprise the terminal repeats of said parvovirus and at least one transcription control sequence selected from the group consisting of a transcription control sequence that regulates the expression of autonomous parvovirus nonstructural polypeptide genes and a transcription control sequence that regulates the expression of autonomous parvovirus structural polypeptide genes.

Claim 25

Dependent Claim 32 words

The vector of claim 1, wherein said heterologous nucleic acid sequences replace autonomous parvovirus sequences from about nucleotide 265 to about nucleotide 4530, wherein said heterologous sequences share substantial homology with LUIII.

Claim 26

Dependent Claim 32 words

The vector of claim 1, wherein said heterologous nucleic acid sequences replace autonomous parvovirus sequences from about nucleotide 145 to about nucleotide 4677, wherein said heterologeous sequences share substantial homology with LUIII.

Claim 27

Dependent Claim 26 words

The vector of claim 1 wherein introduction of said vector into a host cell effects transient gene transfer of said heterologous coding region into said cell.

Claim 28

Dependent Claim 40 words

The vector of claim 2, wherein said heterologous control element comprises a cancer cell-selective response element, wherein said heterologous coding region encodes a cytotoxic agent, and wherein said vector upon introduction into a host cancer cell inhibits cancer cell growth.

Claim 29

Dependent Claim 14 words

The vector of claim 1, wherein said vector comprises a single stranded DNA molecule.

Claim 30

Dependent Claim 14 words

The vector of claim 1, wherein said vector comprises a double stranded DNA plasmid.

Claim 31

Dependent Claim 18 words

The vector of claim 1, wherein said vector is selected from the group consisting of pGLuLUCΔSV and pTOLuLUC.

Claim 32

Dependent Claim 19 words

The vector of claim 1, wherein said vector self-amplifies when provided with viral non-structural proteins by genetically-transformed host cell.

Claim 33

Dependent Claim 11 words

The vector of claim 1, wherein said vector is self-amplification incompetent.

Claim 34

Dependent Claim 20 words

The vector of claim 1, wherein said vector is self-packaging when provided with vector-packaging proteins by a genetically-transformed host cell.

Claim 35

Dependent Claim 11 words

The vector of claim 1, wherein said vector is self-packaging incompetent.

Claim 36

Dependent Claim 85 words

A recombinant virus particle comprising a recombinant vector packaged in an autonomous marvovirus capsid, said vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 37

Dependent Claim 85 words

A recombinant virus particle comprising a recombinant vector packaged in an autonomous marvovirus capsid, said vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 38

Dependent Claim 89 words

A recombinant virus particle comprising a recombinant vector packaged in an autonomous parvovirus capsid, said vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 39

Dependent Claim 89 words

A recombinant virus particle comprising a recombinant vector packaged in an autonomous parvovirus capsid, said vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 40

Dependent Claim 28 words

The virus particle of claim 36, wherein said heterologous nucleic acid sequence is selected from the group consisting of a heterologous control element and a heterologous coding region.

Claim 41

Dependent Claim 52 words

The virus particle of claim 37, wherein said parvovirus nucleic acid sequences are selected from the group consisting of LuIII parvovirus, minute virus of mice, hamster parvovirus, feline panleukopenia virus, canine parvovirus, porcine parvovirus, latent rat virus, mink enteritis virus, human parvovirus, bovine parvovirus, and Aleutian mink disease parvovirus nucleic acid sequences.

Claim 42

Dependent Claim 18 words

The virus particle of claim 36, wherein said parvovirus nucleic acid sequences comprise a LuIII nucleic acid sequence.

Claim 43

Dependent Claim 49 words

The virus particle of claim 36, wherein said capsid is selected from the group consisting of LuIII parvovirus, minute virus of mice, hamster parvovirus, feline panleukopenia virus, canine parvovirus, porcine parvovirus, latent rat virus, mink enteritis virus, human parvovirus, bovine parvovirus, and Aleutian mink disease parvovirus nucleic acid sequences.

Claim 44

Dependent Claim 33 words

The virus particle of claim 36, wherein said capsid is selected from the group consisting of LuIII parvoVirus, minute virus of mice MVMi, minute virus of mice MVMp, and hamster parvovirus H1 capsids.

Claim 45

Dependent Claim 13 words

The virus particle of claim 36, wherein said capsid comprises a LuIII capsid.

Claim 46

Dependent Claim 34 words

The virus particle of claim 36, wherein said recombinant-vector is pseudotyped such that said vector is packaged in a capsid of a virus species other than the species of said parvovirus nucleic acid sequence.

Claim 47

Dependent Claim 53 words

The virus particle of claim 36, wherein said parvovirus nucleic acid sequences comprise a LuIII nucleic acid sequence and wherein said virus capsid is selected from the group consisting of LuIII parvovirus, minute virus of mice, hamster parvovirus, feline panleukopenia virus, canine parvovirus, porcine parvovirus, latent rat virus, and mink enteritis virus capsids.

Claim 48

Dependent Claim 47 words

The virus particle of claim 36, wherein said parvovirus nucleic acid sequences comprise a LuIII nucleic acid sequence and wherein said virus capsid is selected from the group consisting of LuIII parvovirus, minute virus of mice MVMi, minute virus of mice MVMp, and hamster parvovirus H1 capsids.

Claim 49

Dependent Claim 28 words

The virus particle of claim 36, wherein infection of said virus particle into a host cell effects transient gene transfer of said heterologous coding region into said cell.

Claim 50

Dependent Claim 42 words

The virus particle of claim 37, wherein said heterologous control element comprises a cancer cell-selective response element, wherein said heterologous coding region encodes a cytotoxic agent, and wherein infection of said virus particle into a host cancer cell inhibits cancer cell growth.

Claim 51

Dependent Claim 77 words

A gene delivery vehicle comprising a recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 52

Dependent Claim 77 words

A gene delivery vehicle comprising a recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 53

Dependent Claim 31 words

The virus particle of claim 36, wherein said particle exhibits characteristics of an autonomous parvovirus, said characteristics comprising high stability, lack of integration, high titer, and maintenance of infectivity upon concentration.

Claim 54

Dependent Claim 81 words

A gene delivery vehicle comprising a recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeat, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 55

Dependent Claim 81 words

A gene delivery vehicle comprising a recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeat, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 56

Dependent Claim 32 words

The gene delivery vehicle of claim 50, wherein said vector is packaged in an autonomous parvovirus capsid to form a recombinant virus particle effective to deliver said vector to said host cell.

Claim 57

Dependent Claim 21 words

The gene delivery vehicle of claim 51, wherein said capsid targets said virus particle to a selected population of host cells.

Claim 58

Dependent Claim 24 words

The gene delivery vehicle of claim 50, wherein said vector is attached to a carrier effective to deliver said vector to said host cell.

Claim 59

Dependent Claim 20 words

The gene delivery vehicle of claim 53, wherein said carrier is selected from the group consisting of liposomes and viruses.

Claim 60

Dependent Claim 49 words

The gene delivery vehicle of claim 52, wherein said heterologous nucleic acid sequence comprises a control element which is operably linked to a coding region, which control element is selectively functional in a particular population of cells and selectively directs expression of said coding region in said cell population.

Claim 61

Dependent Claim 23 words

The gene delivery vehicle of claim 50, wherein said heterologous nucleic acid sequence encodes an RNA or protein for treating said host cell.

Claim 62

Dependent Claim 72 words

The gene delivery vehicle of claim 50, wherein said vehicle upon introduction into said host cell is capable of substantially destroying a selected population of host cells, said heterologous nucleic acid sequence comprising a heterologous response element that is selectively expressed by said cell population, said response element being operatively linked to a promoter and to a coding region capable of encoding a compound that is substantially cytotoxic to said cell population.

Claim 63

Dependent Claim 29 words

The gene delivery vehicle of claim 57, wherein said compound is selected from the group consisting of a diphtheria toxin, an autonomous parvovirus NS1 protein, and HSV thymidine kinase.

Claim 64

Dependent Claim 82 words

A recombinant nucleic acid comprising nucleic acid sequences of an autonomous parvovirus joined to a heterologous nucleic acid sequence comprising a heterologous control element or heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said recombinant nucleic acid is in a non-integrating form when transferred into a cell.

Claim 65

Dependent Claim 82 words

A recombinant nucleic acid comprising nucleic acid sequences of an autonomous parvovirus joined to a heterologous nucleic acid sequence comprising a heterologous control element or heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said recombinant nucleic acid is in a non-integrating form when transferred into a cell.

Claim 66

Dependent Claim 24 words

The gene delivery vehicle of claim 50, wherein said heterologous nucleic acid sequence restores the function of a defective gene in said host cell.

Claim 67

Dependent Claim 88 words

A recombinant nucleic acid comprising nucleic acid sequences of an autonomous parvovirus joined to a heterologous nucleic acid sequence comprising a heterologous control element or heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said recombinant nucleic acid being in a non-integrating form within a cell after in vitro transfer of said recombinant nucleic acid.

Claim 68

Dependent Claim 88 words

A recombinant nucleic acid comprising nucleic acid sequences of an autonomous parvovirus joined to a heterologous nucleic acid sequence comprising a heterologous control element or heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said recombinant nucleic acid being in a non-integrating form within a cell after in vitro transfer of said recombinant nucleic acid.

Claim 69

Dependent Claim 25 words

The recombinant nucleic acid of claim 61, wherein said heterologous nucleic acid sequence comprises a heterologous control element operatively linked to a heterologous coding region.

Claim 70

Dependent Claim 6 words

The autonomous parvovirus helper construct pSVLu.

Claim 71

Dependent Claim 6 words

The autonomous parvovirus helper construct pSVLu.

Claim 72

Dependent Claim 89 words

A non-integrating vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, the expression of which is regulated by a control element, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said autonomous parvovirus nucleic acid sequences being devoid of nucleic acid sequences encoding either structural or nonstructural autonomous parvovirus polypeptides.

Claim 73

Dependent Claim 89 words

A non-integrating vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, the expression of which is regulated by a control element, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said autonomous parvovirus nucleic acid sequences being devoid of nucleic acid sequences encoding either structural or nonstructural autonomous parvovirus polypeptides.

Claim 74

Dependent Claim 20 words

The vector of claim 64, wherein said vector is packaged within an autonomous parvovirus capsid that target selected cell types.

Claim 75

Dependent Claim 24 words

The vector of claim 64, wherein said vector is capable of effecting transient expression of said heterologous nucleic acid sequence in a host cell.

Claim 76

Independent Claim 107 words

A method for transferring a heterologous nucleic acid sequence into a host cell in vitro comprising introducing into said cell a recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence selected from the group consisting of a heterologous control element and a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 77

Independent Claim 107 words

A method for transferring a heterologous nucleic acid sequence into a host cell in vitro comprising introducing into said cell a recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence selected from the group consisting of a heterologous control element and a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 78

Dependent Claim 18 words

The vector of claim 67, wherein expression of said cytotoxic agent is sufficient to destroy selected cell types.

Claim 79

Independent Claim 111 words

A method for transferring a heterologous nucleic acid sequence into a host cell in vitro comprising introducing into said cell a recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence selected from the group consisting of a heterologous control element and a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 80

Independent Claim 111 words

A method for transferring a heterologous nucleic acid sequence into a host cell in vitro comprising introducing into said cell a recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence selected from the group consisting of a heterologous control element and a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 81

Independent Claim 119 words

A method for transferring a heterologous nucleic acid sequence into a cell in vitro comprising infecting said cell with a recombinant virus particle comprising a recombinant vector packaged in an autonomous parvovirus capsid, said vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence selected from the group consisting of a heterologous control element and a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 82

Independent Claim 119 words

A method for transferring a heterologous nucleic acid sequence into a cell in vitro comprising infecting said cell with a recombinant virus particle comprising a recombinant vector packaged in an autonomous parvovirus capsid, said vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence selected from the group consisting of a heterologous control element and a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 83

Independent Claim 128 words

A method for substantially destroying a selected population of cells comprising administering to an in vitro cell population at least one recombinant vector comprising autonomous parvovirus nucleic acid sequences joined to at least one heterologous nucleic acid sequence having a heterologous response element that is selectively functional in said cell population, said response element being operably linked to a promoter and to a coding region encoding a compound that is substantially cytotoxic to said cell population, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 84

Independent Claim 128 words

A method for substantially destroying a selected population of cells comprising administering to an in vitro cell population at least one recombinant vector comprising autonomous parvovirus nucleic acid sequences joined to at least one heterologous nucleic acid sequence having a heterologous response element that is selectively functional in said cell population, said response element being operably linked to a promoter and to a coding region encoding a compound that is substantially cytotoxic to said cell population, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, wherein said vector is in a non-integrating form when transferred into a cell.

Claim 85

Independent Claim 123 words

A method for transferring a heterologous nucleic acid sequence into a cell in vitro comprising infecting said cell with a recombinant virus particle comprising a recombinant vector packaged in an autonomous parvovirus capsid, said vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence selected from the group consisting of a heterologous control element and a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 86

Independent Claim 123 words

A method for transferring a heterologous nucleic acid sequence into a cell in vitro comprising infecting said cell with a recombinant virus particle comprising a recombinant vector packaged in an autonomous parvovirus capsid, said vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence selected from the group consisting of a heterologous control element and a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 87

Independent Claim 132 words

A method for substantially destroying a selected population of cells comprising administering to an in vitro cell population at least one recombinant vector comprising autonomous parvovirus nucleic acid sequences joined to at least one heterologous nucleic acid sequence having a heterologous response element that is selectively functional in said cell population, said response element being operably linked to a promoter and to a coding region encoding a compound that is substantially cytotoxic to said cell population, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 88

Independent Claim 132 words

A method for substantially destroying a selected population of cells comprising administering to an in vitro cell population at least one recombinant vector comprising autonomous parvovirus nucleic acid sequences joined to at least one heterologous nucleic acid sequence having a heterologous response element that is selectively functional in said cell population, said response element being operably linked to a promoter and to a coding region encoding a compound that is substantially cytotoxic to said cell population, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats, said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 89

Dependent Claim 22 words

The method of claim 73, wherein said coding region encodes an antisense RNA, a ribozyme, an RNA-based drug, or a cytotoxic protein.

Claim 90

Dependent Claim 21 words

The method of claim 73, wherein said selected population of cells comprise cancer cells or cells infected with an infectious agent.

Claim 91

Independent Claim 151 words

A method for producing a recombinant virus particle useful in the delivery of a gone to a targeted cell, comprising: (a) co-transfecting a host cell in vitro with a recombinant non-integrating vector comprising acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence and with a helper construct that effects at least one function selected from the group consisting of amplification of said vector and packaging of said vector in a parvovirus capsid, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats; and (b) culturing said transfected host cell in an effective medium to produce a recombinant virus particle said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 92

Independent Claim 151 words

A method for producing a recombinant virus particle useful in the delivery of a gone to a targeted cell, comprising: (a) co-transfecting a host cell in vitro with a recombinant non-integrating vector comprising acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence and with a helper construct that effects at least one function selected from the group consisting of amplification of said vector and packaging of said vector in a parvovirus capsid, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats; and (b) culturing said transfected host cell in an effective medium to produce a recombinant virus particle said vector being in a non-integrating form within a cell after in vitro transfer of said vector.

Claim 93

Dependent Claim 11 words

The method of claim 76 wherein said helper construct is pSVLu.

Claim 94

Independent Claim 104 words

A method for producing a heterologous product selected from the group consisting of RNA products and protein products comprising: (a) transfecting a host cell in vitro with a recombinant non-integrating vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence encoding said product, said autonomous parvovirus nucleic acid sequences comprising functional left and right inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats; and (b) culturing said transfected host cell in an effective medium to produce said product.

Claim 95

Independent Claim 104 words

A method for producing a heterologous product selected from the group consisting of RNA products and protein products comprising: (a) transfecting a host cell in vitro with a recombinant non-integrating vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence encoding said product, said autonomous parvovirus nucleic acid sequences comprising functional left and right inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats; and (b) culturing said transfected host cell in an effective medium to produce said product.

Claim 96

Dependent Claim 22 words

The method of claim 78, wherein said host cell is further transfected with a helper construct that effects replication of said vector.

Claim 97

Dependent Claim 59 words

A recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats.

Claim 98

Dependent Claim 59 words

A recombinant vector comprising nucleic acid sequences of an autonomous parvovirus joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats.

Claim 99

Dependent Claim 70 words

A recombinant virus particle comprising a recombinant vector packaged in an autonomous parvovirus capsid, said vector comprising autonomous parvovirus nucleic acid sequences joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats.

Claim 100

Dependent Claim 70 words

A recombinant virus particle comprising a recombinant vector packaged in an autonomous parvovirus capsid, said vector comprising autonomous parvovirus nucleic acid sequences joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats.

Claim 101

Dependent Claim 62 words

A gene delivery vehicle comprising a recombinant vector comprising autonomous parvovirus nucleic acid sequences joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats.

Claim 102

Dependent Claim 62 words

A gene delivery vehicle comprising a recombinant vector comprising autonomous parvovirus nucleic acid sequences joined to at least one heterologous nucleic acid sequence, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats.

Claim 103

Dependent Claim 66 words

A recombinant nucleic acid comprising autonomous parvovirus nucleic acid sequences joined to a heterologous nucleic acid sequence comprising a heterologous control element or a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats.

Claim 104

Dependent Claim 66 words

A recombinant nucleic acid comprising autonomous parvovirus nucleic acid sequences joined to a heterologous nucleic acid sequence comprising a heterologous control element or a heterologous coding region, said autonomous parvovirus nucleic acid sequences comprising functional left and right end inverted terminal repeats, said heterologous nucleic acid sequence being located between and operably linked to said nucleic acid sequences comprising said left and right inverted terminal repeats.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1999-12-17	2000-06-17	2000-12-17	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	2003-12-17	2004-06-17	2004-12-17	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2007-12-17	2008-06-17	2008-12-17	$7700.00	Window Closed / Lapsed

Autonomous parvovirus gene delivery vehicles and expression vectors

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