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Method for Site-specific Integration of Nucleic Acids and Related Products (S98028.pdf)

Inventors
Frederic Bushman

Applications
Gene Therapy
Useful for facilitating the identification (i.e., location and isolation) of desired genes.

The invention relates to chimeric proteins useful for targeting and integrating donor nucleic acids at specific locations on target nucleic acids. The invention proteins are also useful for facilitating the identification (i.e., location and isolation) of desired genes. Also provided are nucleic acid constructs encoding invention chimeric proteins, recombinant retroviruses comprising such nucleic acid constructs and methods for site specific control of donor nucleic acid integration into target nucleic acid. Recombinant retroviruses of the invention are useful as attenuated viral vaccines or as vectors for gene therapy methods..

References
Proc. Natl. Acad. Sci. USA 91: 9233-9237
Journal of Virology 71: 458-464

Patent Status:

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Modular Assembly Retroviral Vectors and Uses Therefor (S96013.pdf)

Inventors
Fred H. Gage and Steven T. Suhr

Applications
Gene Expression/Gene Therapy
Gene transfer vectors with the capacity for prolonged or modulated transgene expression

This invention relates to novel retroviral vectors containing modified long terminal repeats (LTR) which enable high level and ligand-modulatable expression of a desired gene product, even after prolonged periods of cellular quiescence. These novel vectors overcome proviral transcriptional inactivation which occurs in cultured primary cells that are growth arrested due to environmental constraints such as contact inhibition and/or nutrient starvation. These vectors represent a class of retroviral vectors in which LTR-promoted proviral expression in infected cells may be maintained or increased, even in situations generally considered to be non-permissive for retroviral vectors. This invention can be applied: as gene transfer vectors with the capacity for prolonged or modulated transgene expression for either in vivo or ex vivo gene therapy; as gene transfer vectors for efficient production of transgenic animals; as vectors for efficient gene transfer to developing embryos; and as vectors with inducible high titers..

References
No publications to date

Patent Status:

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Compositions and Methods for Targeting a Polypeptide to the Central Nervous System (S03007.pdf)

Inventors
Inder Verma and Brian Spencer

Applications
Gene Therapy, Drug Development
Identification of modular targeting molecules that can selectively penetrate the blood-brain barrier

There exists a need for a mode or method that allows the passage of a specific therapeutic polypeptide across the blood-brain barrier. This invention is directed to the identification of modular targeting molecules that can selectively penetrate the blood-brain barrier (BBB). These chimeric CNS targeting polypeptides are comprised of a BBB binding domain and a payload polypeptide domain. The targeting molecules can carry and deliver any polypeptide of interest to the central nervous system (CNS). Such CNS targeting polypeptides have the advantage in that they can be administered directly to an individual, or they can be expressed via an encoding nucleic acid by non-target cells, and they will travel to and concentrate in the CNS. This approach will be applicable to lysosomal storage diseases such as Gaucher's disease, Hunter's disease and Fabry Syndrome. In addition the fusion of ligands to facilitate passage of proteins across the blood-brain barrier may be a general method for delivering therapeutic proteins to the CNS for neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease or prion diseases..

References
Published PCT Application WO04/108071

Patent Status:
U.S. Application filed June 5, 2003

License Terms:
Exclusive, Partially Exclusive, Nonexclusive license negotiable

Reference_Number: S03007
Contact: Mike White, Ph.D., CLP o Director, OTM o 858.453.4100 x1703 o mwhite@salk.edu

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Compositions and Methods for Tissue Specific Targeting (S02015B.pdf)

Inventors
Inder Verma, Brian Spencer, Robert Marr

Applications
Lentiviral vectors, Gene Therapy
Lentiviral targeting vectors capable of being targeted to any desired cell or tissue that contains an identifying cell surface marker.

Therapies for certain diseases will require gene delivery to a specific tissue or organ, e.g. diabetes (pancreas), cystic fibrosis (lung), hemophilia (liver or hematopoetic stem cells), hypercholesteremia (liver). Although tissue specific promoters can be used to restrict expression of the gene to a tissue, many of these promoters appear to allow limited gene expression in other tissues while others restrict gene expression to a short time frame after gene delivery. Furthermore, many tissues and cells will still be infected with the virus if the transgene is not expressed. Salk scientists have developed a novel approach to selectively and rationally target the lentiviral vector to tissues or organs based on receptor expression. This invention is directed to lentiviral targeting vectors that are capable of being targeted to essentially any desired cell or tissue that contains an identifying cell surface marker. These lentiviral vectors separate the cell binding and attachment functions from the membrane fusion functions of a lentiviral envelope polypeptide. Accordingly, the vectors are advantageous in that the infection specificity and the transduction activities constitute modular components of the viral vector. By separating these activities into targeting and fusogenic polypeptides, the inherent constraints associated with linked activities when modification is necessary or desired are circumvented because each activity can be separately manipulated. This approach may be useful for directing gene therapy treatment to specific tissues and organs. In addition, gene delivery to the whole brain to treat global neuronal degenerative disorders could best be accomplished by targeting the viral vector for transport across the blood brain barrier and thus widespread distribution over the whole brain..

References
No publications to date

Patent Status:
U.S. Application Published as 2005-0003541

License Terms:
Exclusive, Partially Exclusive, Nonexclusive license negotiable

Reference_Number: S02015B
Contact: Mike White, Ph.D., CLP o Director, OTM o 858.453.4100 x1703 o mwhite@salk.edu

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Methods of Inhibiting Gene Expression by RNA Interference Lentiviral Vectors to Generate Functional Knock-outs in Cells, Tissues and Animals (S02014.pdf)

Inventors
Inder Verma, Gustavo Tiscornia, Oded Singer

Applications
Gene silencing, Gene Therapy, Transgenesis, Functional Genomics
Use of lentiviral vectors expressing small interfering RNAs (siRNAs) to knock down the expression of specific genes in vitro and in vivo

Recently several approaches have been described for generating loss-of-function phenotypes in mammalian systems by using RNAi. All these approaches, however, have limited applications and are not applicable for generating a long-term silencing effect in vivo. To overcome these limitations, scientists at the Salk Institute have designed a lentiviral vector system capable of expressing siRNA in various mammalian systems. This lentivirus system can express integrated siRNA efficiently in a wide variety of cell lines and primary cells both in vitro and in vivo. More importantly, the recombinant viruses can be used to infect fertilized eggs in vitro to generate transgenic mice where specific gene(s) function(s) is compromised. The ease and convenience of deleting gene function combined with the ability to make transgenic animals with the appropriate mutant gene will be very useful for proof of concept for gene therapy experiments. The transgenic mice containing the actual mutant gene will also be useful to study immunological consequences of the transgene introduced by gene delivery vectors and allow for the development of gene knockouts in species other than mice. Lastly, this invention could be helpful for gene therapy of genetic disease due to gain of function such as Huntington's disease..

References
PNAS, Vol. 100, No. 4, 1844-1848 (February 2003)

Patent Status:
U.S. Application Published as 2005-0234504

License Terms:
For Internal Research Purposes Only - Exclusive, Partially Exclusive, Nonexclusive license negotiable

Reference_Number: S02014
Contact: Mike White, Ph.D., CLP o Director, OTM o 858.453.4100 x1703 o mwhite@salk.edu

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Retroviral Packaging Cell Line (S97022A.pdf)

Inventors
Inder Verma, Tal Kafri, Frederic Bushman, Mark Hansen

Applications
Gene Therapy
Produces high titers of pseudotyped retroviral particles capable of transducing nondividing cells.

This invention describes a packaging cell line that produces retroviral particles, especially pseudotyped particles at a higher titer than conventional packaging cell lines. An advantage of the invention is that packaging cell lines can be produced that contain an envelope-encoding nucleotide sequence stably integrated in the cell's genome, which sequence can be inducibly expressed, thus allowing generation of packaging cell lines capable of expressing an envelope protein that is otherwise toxic to the host cell. Also, the packaging cell lines do not have the potential to produce replication competent retroviruses. The inventors have developed HIV vectors that can transduce nondividing cells. The HIV vector was pseudotyped with the vesicular stomatitis virus G glycoprotein to ensure a broad host range and facilitate concentration of virus to high titers. HIV vectors mediate efficient and stable transduction of post-mitotic cells in brain, liver, muscle and retina. Recently, the inventors were able to show that a lentiviral vector based on HIV was able to transduce human CD34+ cells capable of stable, long-term reconstitution of non obese diabetic/severe combined immunodeficient (NOD/SCID) mice. This opens the way for human gene therapy using human hematopoietic stem cells..

References
Science Vol. 283, p. 682-686 (1999)

Patent Status:
U.S. Patent Number 6,727,058 issued April 27, 2004
Foreigns pending

License Terms:
Nonexclusive license negotiable

Reference_Number: S97022A
Contact: Mike White, Ph.D., CLP o Director, OTM o 858.453.4100 x1703 o mwhite@salk.edu

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Somatic Cell Gene Therapy (S94004.pdf)

Inventors
Inder Verma and Daniel St. Louis

Applications
Gene Therapy
A gene therapy method of implanting transduced fibroblasts in the loose connective tissue of the skin

This invention is directed to fibroblast cells which are transduced so that they express a "replacement" gene of interest. These transduced fibroblasts are preferably fixed in vitro in an extracellular matrix, and then implanted in the loose connective tissue of the skin of an individual or animal to be treated. Because the fibroblasts are implanted in a highly vascularized compartment of the skin, the transduced cells, and thus their "replacement" gene products, have direct access to the circulatory system. As a result the needed replacement gene products can easily and efficiently be distributed to other parts of the body. When the gene therapy is no longer needed, the implanted fibroblasts can be conveniently removed..

References
Proc. Natl. Acad. Sci. USA, vol. 85:3150-3154 (1988)

Patent Status: