| Lentiviral ltr-deleted vector -> Monitor Keywords |
|
|
 
Lentiviral ltr-deleted vectorRelated Patent Categories: Chemistry: Molecular Biology And Microbiology, Process Of Mutation, Cell Fusion, Or Genetic Modification, Introduction Of A Polynucleotide Molecule Into Or Rearrangement Of Nucleic Acid Within An Animal Cell, The Polynucleotide Is Encapsidated Within A Virus Or Viral CoatLentiviral ltr-deleted vector description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20060177934, Lentiviral ltr-deleted vector. Brief Patent Description - Full Patent Description - Patent Application Claims
RELATED APPLICATIONS [0001] This application is a continuation-in-part of U.S. application Ser. No. 09/254,832, filed on Jun. 21, 1999, as the national phase application of International application Serial No. PCT/GB97/02969, filed on Oct. 28, 1997 and claiming priority to UK application Serial No. GB 9622500.8, filed on Oct. 29, 1996. This application makes reference to U.S. Pat. No. 6,235,522, filed on Apr. 5, 1999 as the national phase application of International application Serial No. PCT/GB97/02858, filed on Oct. 17, 1997 and claiming priority to UK application Serial No. GB 9621680. This application also makes reference to: U.S. Pat. No. 6,096,538, filed on Nov. 19, 1997, U.S. Pat. No. 6,132,731, filed on Oct. 8, 1997, U.S. Pat. No. 6,168,916, filed on Oct. 21, 1998, U.S. Pat. No. 6,312,682, filed on Dec. 28, 1998, U.S. Pat. No. 6,312,683, filed on Jan. 27, 1999, U.S. application Ser. No. 09/533,276, filed on Mar. 22, 2000, U.S. application Ser. No. 09/533,295, filed on Mar. 22, 2000, U.S. application Ser. No. 09/552,950, filed on Apr. 20, 2000, U.S. application Ser. No. 09/860,996, filed on May 18, 2001, U.S. application Ser. No. 09/867,947, filed on May 29, 2001, U.S. application Ser. No. 09/915,169, filed on Jul. 25, 2001, U.S. application Ser. No. 10/001,220, filed on Nov. 15, 2001, U.S. application Ser. No. 10/002,598, filed on Nov. 15, 2001, and U.S. application Ser. No. 10/008,610, filed on Nov. 8, 2001. [0002] Each document cited or referenced in each of the foregoing applications, and any manufacturer's instructions or catalogues for any products cited or mentioned in each of the foregoing applications and in any of the cited documents, are hereby incorporated herein by reference. Furthermore, all documents cited in this text, all documents cited or referenced in documents cited in this text, and any manufacturer's instructions or catalogues fanny products cited or mentioned in this text or in any document incorporated into this text, are incorporated herein by reference. Documents incorporated by reference into this text or any teachings therein can be used in the practice of this invention. Documents incorporated by reference into this text are not admitted to be prior art. FIELD OF THE INVENTION [0003] This invention relates to lentiviral long terminal repeat (LTR)-deleted vectors. The invention also relates to lentiviral LTR-deleted vectors carrying nucleotide sequences of interest, and to their use in transferring genetic material to non-dividing or slowly dividing BACKGROUND OF THE INVENTION [0004] Amongst nucleic acid transfer systems, retroviral vectors hold substantial promise for gene therapy and other applications in which transfer of genetic material is desirable. These systems can transfer genes efficiently, and new vectors are emerging that are particularly useful for gene delivery to brain cells (Naldini et al., 1996 Science 272, 263). [0005] There has been considerable interest in the development of retroviral vector systems based on lentiviruses, a small subgroup of the retroviruses. This interest arises firstly from the notion of using HIV-based vectors to target anti-HIV therapeutic genes to HIV susceptible cells and secondly from the prediction that, because lentiviruses are able to infect non-dividing cells (Lewis & Emerman 1993 J. Virol. 68, 510), vector systems based on these viruses are able to transduce non-dividing cells (e.g. Vile & Russel 1995 Brit. Med. Bull. 51, 12). Vector systems based on HIV have been produced (Buchschacher & Panganiban 1992 J. Virol. 66, 2731) and have been used to transduce CD4+ cells and non-diving cells (Naldini et al., 1996 Science 272, 263). However, in general, nucleic acid transfer efficiencies are not as high as with comparable murine retrovirus vector systems. [0006] The HIV-based vectors produced to date result in an integrated provirus in the transduced cell that has HIV LTRs at its ends. This limits the use of these vectors as the LTRs have to be used as expression signals for any inserted gene unless an internal promoter is used. The use of internal promoters has significant disadvantages. For example, the presence of internal promoters can affect the transduction titres obtainable from a packaging cell line and the stability of the integrated vector. [0007] Also, HIV and other lentiviral LTRs have virus-specific requirements for nucleic acid expression. For example, the HIV LTR is not active in the absence of the viral Tat protein (Cullen 1995 AIDS 9, S19). It is desirable, therefore, to modify or delete the LTRs in such a way as to change the requirements for nucleic acid expression. In particular, tissue specific gene expression signals may be required for some gene therapy applications. In addition, signals that respond to exogenous signals may be necessary. In murine retroviruses this is often achieved simply by replacing the enhancer-like elements in the U3 region of the murine lentiviral (MLV) LTR by enhancers that respond to the desired signals. This has not been feasible with viruses such as HIV because within the U3 and R regions of their LTRs are sequences, known as IST and TAR, which may inhibit gene expression and may or may not be responsive to Tat protein when heterologous, perhaps tissue specific, control sequences are inserted in the U3 region (Cullen 1995 AIDS 9, S19; Alonso et al., 1994 J. Virol. 68, 6505; Ratnasabapathy et al., 1990 4, 2061; Sengupta et al., 1996 PNAS 87, 7492; Parkin et al., 1988 EMBO. J 7, 2831). Even if the signals are responsive, it is undesirable to have to supply Tat as it further complicates the system and Tat has some properties of oncoproteins (Vogel et al., 1988 Nature 335, 606). [0008] Parkinson's disease (PD) is a common neurodegenerative disorder that afflicts the growing population of elderly people. Patients display tremor, cogwheel rigidity and impairment of movement. It is generally thought to be an acquired rather than inherited disease in which environmental toxins, metabolic disorders, infectious agents and normal aging have all been implicated. PD is associated with the degeneration of nigrostriatal neurons which have their soma located in the substantia nigra. They send axonal projections to the basal ganglia and they use dopamine as their neurotransmitter. Some features of the disease can be controlled by the administration of L-DOPA, the metabolic precursor to dopamine, which diffuses across the blood brain barrier more effectively than dopamine itself. Unfortunately as the disease progresses the side effects of this treatment become unacceptable. [0009] PD is an ideal candidate for gene therapy for several reasons. The clinical efficacy of systemic administration of L-DOPA suggests that restoration of neuronal circuitry is not essential for disease management. Therefore genetic manipulation of brain cells to provide local production of L-DOPA from tyrosine may be a realistic strategy for treatment. The biosynthesis of L-DOPA from tyrosine involves a single step suggesting that provision of tyrosine hydroxylase (TH) by genetic means may be sufficient and some success has been achieved using this strategy in small animals and in cell culture (Kaplitt et al., 1994 Nature Genetics 8, 148; During et al., 1994 Science 266, 1399; Horellou et al., 1994 Neuroreport 6, 49; Owens et al., 1991 J. Neurochem. 56, 1030). However, if one is to use local endogenous brain cells as L-DOPA factories for the treatment of PD in man it is likely that high levels of L-DOPA will be required to effect a treatment. These high levels must be efficiently converted to dopamine as the necessary neurotransmitter and primary therapeutic agent. It is likely therefore that it will be necessary not only to supply tyrosine hydroxylase but also DOPA decarboxylase (DD), the enzyme that converts L-DOPA to dopamine. This means that in a gene therapy strategy the genes for both of these enzymes will be required. However, it is clear from the literature that retroviral vectors achieve the highest titres and most potent gene expression properties if they are kept genetically simple (PCT/GB96/01230; Bowtell et al., 1988 J. Virol. 62, 2464; Correll et al., 1994 Blood 84, 1812; Emerman and Temin 1984 Cell 39, 459; Ghattas et al., 1991 Mol. CeII. Biol. 11, 5848; Hantzopoulos et al., 1989 PNAS 86, 3519; Hatzoglou et al., 1991 J. BioL Chem 266, 8416; Hatzoglou et al., 1988 J. BioI. Chem 263, 17798; Li et al., 1992 Hum. Gen. Ther. 3, 381; McLachlin et al., 1993 Virol. 195, 1; Overell et al., 1988 MoI. Cell Biol. 8, 1803; Scharfman et al., 1991 PNAS 88, 4626; Vile et al., 1994 Gene Ther 1, 307; Xu et al., 1989 Virol. 171, 331; Yee et al., 1987 PNAS 84, 5197). This means only using a single transcription unit within the vector genome and orchestrating appropriate nucleic acid expression from sequences within the 5' LTR. The need to express two enzymes from a single retroviral vector would require the use of an internal ribosome entry site (IRES) to initiate translation of the second coding sequence in a poly-cistronic message (Adam et al. 1991 J. Virol. 65, 4985). However, the efficiency of an IRES is often low and tissue dependent making this strategy undesirable when one is seeking to maximise the efficiency of metabolic conversion of tyrosine through to dopamine. The present invention addresses these problems. SUMMARY OF THE INVENTION [0010] The present invention provides in one aspect a lentiviral vector capable of transducing a non-dividing or slowly-dividing cell, said vector comprising a lentiviral LTR-deleted vector. The vector can further comprise a nucleotide sequence encoding a polypeptide or protein of interest (POI), e.g., at least one nucleotide sequence of interest (NOI) encoding at least one POI. Advantageously, the NOI is operably linked to a promoter. If there is more than one NOI, there can be one promoter for driving expression, or a promoter for each NOI for driving expression. Thus, one or more NOI can be operably linked to one or more NOI. The vector can comprise a polynucleotide sequence, which encodes two or more POI, e.g., therapeutic POI, operably linked to a promoter, and the polynucleotide can encode a fusion POI. The invention thus can provide a way of expressing two therapeutic NOI from a single "chimeric" gene or polynucleotide. The vector may be for example an expression vector such as a plasmid, or it may be a retroviral vector particle comprising an RNA genome containing the nucleotide sequences as described herein. [0011] In another aspect, the invention provides a method for producing a POI in a non-dividing or slowly-dividing cell, comprising transducing the cell with a lentiviral LTR-deleted vector and expressing the POI in the cell. In a preferred embodiment, the non-dividing cell is a neuron. [0012] There are many uses for in vitro expressed POI. For instance, depending on the nature of the POI the in vitro expressed POI can represent a protein that is purer than if the POI was isolated from its native environment, as it would be free from contaminants from that environment. Thus, such POI can be used in assays, to generate antibodies, e.g. for use in assays, as antigens or epitopes in immunological compositions, and as active agents in therapeutic, pharmaceutical or veterinary compositions, inter alia. [0013] The invention further provides a target cell in vitro comprising a lentiviral LTR-deleted vector. In yet further aspects, the invention provides a DNA construct encoding the RNA genome for the retroviral vector particle; and a retroviral vector production system comprising a set of nucleic acid sequences encoding the components of the retroviral vector particle. [0014] The invention further provides the use of retroviral vectors carrying the chimeric gene described herein, in gene therapy and in the preparation of a medicament for gene therapy; and a method of performing gene therapy on a target cell, which method comprises transducing the target cell with a lentiviral LTR-deleted vector comprising a nucleotide sequence encoding a POI, thus delivering the nucleotide sequence to the target cell. The invention further provides transduced target cells resulting from these methods and uses. The invention thus provides a gene delivery system for use in medicine. [0015] The term "comprising" in this disclosure can mean "including" or can have the meaning commonly given to the term "comprising" in U.S. Patent Law. [0016] Other aspects of the invention are described in or are obvious from (and within the ambit of the invention) the following disclosure. BRIEF DESCRIPTION OF DRAWINGS [0017] The following Detailed Description, given by way of example; but not intended to limit the invention to specific embodiments described, may be understood in conjunction with the accompanying drawings, incorporated herein by reference, in which: [0018] FIG. 1 shows a general scheme for Lentiviral LTR-deleted (LLD) vectors which may be used with the present invention and which are employed in the Examples. [0019] FIG. 2 shows a generalised HIV-based LLD vector genome as described in the Examples; Superscript H=HIV-derived sequence (could be from any lentivirus); Superscript M=MLV-derived sequence; 1V=Packaging site (including gag region); PBS=Second strand priming site; INTERNAL=Region containing genes, selectable markers, other promoters or RNA handling systems such as HIV RRE and Rev coding sequences. Continue reading about Lentiviral ltr-deleted vector... Full patent description for Lentiviral ltr-deleted vector Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Lentiviral ltr-deleted vector patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. Each week you receive an email with patent applications related to your keywords. Start now! - Receive info on patent apps like Lentiviral ltr-deleted vector or other areas of interest. ### Previous Patent Application: Expression of apoa-1 and variants thereof using spliceosome mediated rna trans-splicing Next Patent Application: Modified freeze fracture direct imaging apparatus and technique Industry Class: Chemistry: molecular biology and microbiology ### FreshPatents.com Support Thank you for viewing the Lentiviral ltr-deleted vector patent info. IP-related news and info Results in 0.49468 seconds Other interesting Feshpatents.com categories: Computers: Graphics , I/O , Processors , Dyn. Storage , Static Storage , Printers 174 |
 * Protect your Inventions * US Patent Office filing 
PATENT INFO |
|
