Involving General Or Homologous Recombination (e.g., Gene Targeting, Etc.) Patents (Class 435/463)
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Publication number: 20020025578Abstract: A novel method is provided for delivering nucleic acid molecules to the cells of an organism by pulse voltage delivery. The method involves the combination of formulated nucleic acid molecules with devices for injecting the molecules by pulse voltage or an electrical field. Disclosed are compositions and methods for enhancing the administration to and uptake of nucleic acids in a mammal. The methods disclosed provide an increased transfection and/or gene delivery efficiency by enhancing the uptake of formulated nucleic acid molecules by applying an electrical field which destabilizes the cellular membrane thereby opening pores or passageways which allow extracellular material to be introduced to the cell. Also disclosed are examples which demonstrate that the combination of formulated nucleic acid molecules and pulse voltage injection methods results in immune responses which are superior to those obtained by conventional means of delivery.Type: ApplicationFiled: July 10, 2001Publication date: February 28, 2002Applicant: Valentis, Inc.Inventors: Fiona MacLaughlin, Shulin Li, Yuhua Li, Karel Petrak
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Patent number: 6350932Abstract: A transgenic mouse containing disruptions in both vitamin D alleles and lacking vitamin D receptor activity is described. The transgenic mouse displays perioral and periorbital alopecia, hypocalcemia, hypophosphatemia, and bone demineralization. The transgenic mouse is useful for screening treatments for a number of conditions associated with vitamin D receptor related disorders including skin disorders, immune system disorders, and proliferative disorders.Type: GrantFiled: August 28, 1998Date of Patent: February 26, 2002Assignee: The General Hospital CorporationInventors: Marie Demay, Yan Chun Li
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Patent number: 6351690Abstract: A system for performing antiviral drug susceptibility and resistance testing is automated using software and robotics. The system includes a transfection apparatus, an infection apparatus and a plate reading apparatus. One or more of the apparatuses may be automated.Type: GrantFiled: January 21, 2000Date of Patent: February 26, 2002Assignee: Virologic, Inc.Inventor: Steven J. Lenz
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Patent number: 6346414Abstract: A transposition assembly for the transfer of a DNA fragment of interest into the ribosomal nuclear DNA of an eukaryotic cell. An insertion means, an eukaryotic cell and a pharmaceutical composition comprising said transposition assembly, as well as a method for the in vitro transfer of said DNA fragment, are also disclosed.Type: GrantFiled: October 16, 1995Date of Patent: February 12, 2002Assignee: Transgene S.A.Inventor: Eric Jacobs
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Patent number: 6326206Abstract: The present invention relates to a method for in vivo recombination of homologous DNA sequences. The method is a forced artificial evolution resulting in a DNA sequence encoding a polypeptide having an advantageous property.Type: GrantFiled: June 10, 1999Date of Patent: December 4, 2001Assignee: Novozymes A/SInventors: Mads Eskelund Bjornvad, Michael Dolberg Rasmussen, Per Lina Jorgensen, Torben Vedel Borchert, Stanislas Dusko Ehrlich
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Publication number: 20010046707Abstract: The invention provides a number of strategies for transferring and/or evolving gene(s) associated with cellular DNA uptake so that they confer or enhance DNA-uptake capacity of a recipient cell. Evolution is achieved by recursive cycles of recombination and screening/selection. One such strategy entails evolving genes that confer competence in one species to confer either greater competence in that species, or comparable or greater competence in a second species. Another strategy entails evolving genes for use as components of cloning vector to confer enhanced uptake of the vector. Other strategies entail evolving viral receptors, viruses, and genes that mediate conjugal transfer.Type: ApplicationFiled: November 1, 1999Publication date: November 29, 2001Inventor: WILLEM P. C. STEMMER
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Patent number: 6303379Abstract: The present invention relates to transfected primary and secondary somatic cells of vertebrate origin, particularly mammalian origin, transfected with exogenous genetic material (DNA) which encodes a desired (e.g., a therapeutic) product or is itself a desired (e.g., therapeutic) product, methods by which primary and secondary cells are transfected to include exogenous genetic material, methods of producing clonal cell strains or heterogenous cell strains, methods of gene therapy in which the transfected primary or secondary cells are used, and methods of producing antibodies using the transfected primary or secondary cells.Type: GrantFiled: July 16, 1999Date of Patent: October 16, 2001Assignee: Transkaryotic Therapies, Inc.Inventors: Richard F Selden, Douglas Treco, Michael W. Heartlein
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Patent number: 6297029Abstract: A mammalian chromosome is produced according to the method comprising the steps of: introducing a DNA construct comprising a mammalian telomere and a centromere into a mammalian cell, wherein the centromere has a DNA sequence containing a plurality of copies of the CENP-B box sequence consisting of: 5′-NTTCGNNNNANNCGGGN-3′, wherein N is any one of A, T, C and G.Type: GrantFiled: September 28, 1999Date of Patent: October 2, 2001Inventors: Tsuneko Okazaki, Hiroshi Masumoto, Masashi Ikeno, Howard J. Cooke, Brenda Rose Grimes
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Patent number: 6294385Abstract: A method for making insertional mutations at random or quasi-random locations in the chromosomal or extra-chromosomal nucleic acid of a target cell includes the step of combining, in the target cell, cellular nucleic acid with a synaptic complex that comprises (a) a transposase protein and (b) a polynucleotide that comprises (1) a pair of nucleotide sequences adapted for operably interacting with Tn5 transposase and (2) a transposable nucleotide sequence therebetween, under conditions that mediate transpositions into the cellular DNA. In the method, the synaptic complex is formed in vitro under conditions that disfavor or prevent the synaptic complexes from undergoing productive transposition.Type: GrantFiled: August 10, 2000Date of Patent: September 25, 2001Assignee: Wisconsin Alumni Research FoundationInventors: Igor Y. Goryshin, William S. Reznikoff
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Patent number: 6291165Abstract: The present invention relates to a new method of shuffling especially heterologous polynucleotide sequences, screening and/or selection of new recombinant proteins resulting therefrom having a desired biological activity, and especially to production and identification of novel proteases exhibiting desired properties.Type: GrantFiled: March 17, 1998Date of Patent: September 18, 2001Assignee: Novo Nordisk A/SInventors: Torben Vedel Borchert, Titus Kretzschmar, Joel R. Cherry
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Patent number: 6284541Abstract: The invention concerns a method for the introduction of a foreign DNA into the genome of a target cell by homologous recombination as well as for the homologous recombination of suitable DNA constructs.Type: GrantFiled: June 9, 2000Date of Patent: September 4, 2001Assignee: Roche Diagnostics GmbHInventors: Johannes Auer, Raimund Sprenger, Konrad Honold
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Patent number: 6278040Abstract: In accordance with the present invention, there are provided targeted loss of function mutant mice which express less than endogenous levels of at least one member of the steroid/thyroid superfamily of receptors in at least one specific tissue type. For example, mutations in the RXR&agr; gene in mouse germlines are lethal in the embryonic stage between E13.5 and E16.5 when bred to homozygosity. The major defect responsible for this lethal effect is hypoplastic development of the ventricular chambers of the heart, which is manifest as a grossly thinned ventricular wall with concurrent defects in ventricular septation. This phenotype is identical to a subset of the effects of embryonic vitamin A deficiency, and therefore establishes RXR&agr; as a genetic component of the vitamin A signaling pathway in cardiac morphogenesis.Type: GrantFiled: February 19, 1997Date of Patent: August 21, 2001Assignees: The Salk Institute for Biological Studies, The Regents of the University of Calif.Inventors: Henry M. Sucov, Ronald M. Evans, Kenneth R. Chien
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Patent number: 6270989Abstract: The invention relates to novel human DNA sequences, targeting constructs, and methods for producing novel genes encoding thrombopoietin, DNase I, and &bgr;-interferon by homologous recombination. The targeting constructs comprise at least: a) a targeting sequence; b) a regulatory sequence; c) an exon; and d) a splice-donor site. The targeting constructs, which can undergo homologous recombination with endogenous cellular sequences to generate a novel gene, are introduced into cells to produce homologously recombinant cells. The homologously recombinant cells are then maintained under conditions which will permit transcription of the novel gene and translation of the mRNA produced, resulting in production of either thrombopoietin, DNase I, or &bgr;-interferon.Type: GrantFiled: March 17, 1995Date of Patent: August 7, 2001Assignee: Transkaryotic Therapies, Inc.Inventors: Douglas A. Treco, Michael W. Heartlein, Brian M. Hauge, Richard F Selden
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Patent number: 6271360Abstract: The invention concerns the introduction of predetermined genetic changes in target genes of a living cell by introducing an oligodeoxynucleotide encoding the predetermined change. The oligodeoxynucleotides are effective in mammalian, avian, plant and bacterial cells. Specific end modifications that greatly increase the effectiveness of the oligodeoxynucleotides in bacteria are described. Surprisingly, unmodified oligodeoxynucleotides can be as effective in mammaliancells, including in vivo hepatocytes, as the modified nucleotides and can be as effective or more effective than chimeric oligonucleotides that consist of a mixture of deoxynucleotides and 2′-O-methyl ribonucleotides.Type: GrantFiled: August 27, 1999Date of Patent: August 7, 2001Assignee: ValiGen (US), Inc.Inventors: Richard A. Metz, Bruce L. Frank, Debra M. Walther
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Patent number: 6270996Abstract: An adenovirus E1/E4 expressing packaging cell line is provided, which permits the generation of recombinant adenoviruses deleted in both gene regions. A method for enhancing the efficiency of transduction of a recombinant AAV into a target cell is provided by infecting a target cell with a recombinant AAV comprising a selected transgene under the control of regulatory sequences. The infected cell is contacted with an agent which facilitates the conversion of single stranded recombinant virus to its double stranded form.Type: GrantFiled: May 3, 2000Date of Patent: August 7, 2001Assignee: The Trustees of the University of PennsylvaniaInventors: James M. Wilson, Krishna J. Fisher, Guang-Ping Gao
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Patent number: 6271436Abstract: Disclosed are methods for the isolation of primordial germ cells, culturing these cells to produce primordial germ cell-derived cell lines, methods for transforming both the primordial germ cells and the cultured cell lines, and using these transformed cells and cell lines to generate transgenic animals. The efficiency at which transgenic animals are generated by the present invention is greatly increased, thereby allowing the use of homologous recombination in producing transgenic non-rodent animal species.Type: GrantFiled: October 10, 1997Date of Patent: August 7, 2001Assignee: The Texas A & M University SystemInventors: Jorge A. Piedrahita, Fuller W. Bazer
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Reagents and methods for the screening of compounds useful in the treatment of neurological diseases
Patent number: 6268216Abstract: This invention relates to the gene sequence of a novel transcription factor specific for central 5-HT (serotonergic) neurons. The sequence and products are useful in screening methods for identifying and testing agonists and antagonists of seronergic activity.Type: GrantFiled: July 26, 1999Date of Patent: July 31, 2001Assignee: Case Western Reserve UniversityInventors: Evan Samuel Deneris, Dmitry Viktor Fyodorov, Timothy John Hendricks -
Patent number: 6261551Abstract: An adenovirus E1/E4 expressing packaging cell line is provided, which permits the generation of recombinant adenoviruses deleted in both gene regions. A method for enhancing the efficiency of transduction of a recombinant AAV into a target cell is provided by infecting a target cell with a recombinant AAV comprising a selected transgene under the control of regulatory sequences. The infected cell is contacted with an agent which facilitates the conversion of single stranded recombinant virus to its double stranded form.Type: GrantFiled: December 5, 1997Date of Patent: July 17, 2001Assignee: The Trustees of the University of PennsylvaniaInventors: James M. Wilson, Krishna J. Fisher, Guang-Ping Gao
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Publication number: 20010008026Abstract: The present invention relates to methods and compositions for tissue-restricted gene recombination. In particular, the present invention provides methods and compositions for tissue-restricted gene recombination in post-mitotic cells. The present invention further provides methods for gene recombination in post-mitotic cells comprising the delivery of a Cre recombinase to the target tissue to facilitate recombination in a desired target nucleic acid.Type: ApplicationFiled: June 25, 1998Publication date: July 12, 2001Inventors: MICHAEL D. SCHNEIDER, PAUL OVERBEEK, PETER FRENKEL
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Patent number: 6255113Abstract: The invention relates to methods for targeting an exogenous polynucleotide or exogenous complementary polynucleotide pair to a predetermined endogenous DNA target sequence in a eukaryotic cell by homologous pairing, particularly for altering an endogenous DNA sequence, such as a chromosomal DNA sequence, typically by targeted homologous recombination. In certain embodiments, the invention relates to methods for targeting an exogenous polynucleotide having a linked chemical substituent to a predetermined endogenous DNA sequence in a metabolically active eukaryotic cell, generating a DNA sequence-specific targeting of one or more chemical substituents in an intact nucleus of a metabolically active eukaryotic cell, generally for purposes of altering a predetermined endogenous DNA sequence in the cell.Type: GrantFiled: February 8, 1995Date of Patent: July 3, 2001Assignee: SRI InternationalInventors: David A. Zarling, Elissa P. Sena
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Patent number: 6251677Abstract: The present invention provides a hybrid vector construct which comprises a portion of an adenovirus, 5′ and 3′ ITR sequences from an AAV, and a selected transgene. Also provided is a hybrid virus linked via a polycation conjugate to an AAV rep gene to form a single particle. These trans-infection particles are characterized by high titer transgene delivery to a host cell and the ability to stably integrate the transgene into the host cell chromosome. Also disclosed is the use of the hybrid vectors and viruses to produce large quantities of recombinant AAV.Type: GrantFiled: February 8, 1999Date of Patent: June 26, 2001Assignee: The Trustees of the University of PennsylvaniaInventors: James M. Wilson, William M. Kelley, Krishna J. Fisher
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Patent number: 6245963Abstract: A genetically engineered mouse model that genotypically and phenotypically mimics human patients with spinal muscular atrophy. The genome of the mouse model contains at least one mutation that knockouts the native mouse Smn gene and at least one copy of human SMNC gene that functions in a murine background and compensates for the loss of the functions provided by the Smn gene. The phenotypes of said mouse model can be grouped according to their severity of pathological conditions into three types, paralleling the three types of human spinal muscular atrophy conditions. Said mouse model can be used for studying the pathophysiology of spinal muscular atrophy and for developing and testing existing and new therapeutic and diagnostic methods.Type: GrantFiled: May 25, 2000Date of Patent: June 12, 2001Assignee: Academia SinicaInventors: Hung Li, Hsiu-Mei Hsieh-Li, Jan-Gowth Chang
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Patent number: 6242211Abstract: The present invention relates to a novel drug discovery system for generating molecular diversity. The system provides methods for subjecting the genetic materials from a plurality of species of organisms to homologous or homeologous recombination to create novel genes and metabolic pathways. The recombined genetic materials are cloned to form recombined combinatorial gene expression libraries. Methods for screening such gene expression libraries containing recombined genes and metabolic pathways for novel activities and compounds are also provided.Type: GrantFiled: March 5, 1999Date of Patent: June 5, 2001Assignee: Terragen Discovery, Inc.Inventors: Todd C. Peterson, Paul Brian
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Patent number: 6242218Abstract: An isolated nucleic acid molecule that hybridizes under stringent conditions, or shares at least 80% sequence identity, with a defined genomic region upstream of the coding region of the G-CSF gene, and a DNA construct containing that DNA molecule as a targeting sequence for homologous recombination.Type: GrantFiled: May 5, 1999Date of Patent: June 5, 2001Assignee: Transkaryotic Therapies Inc.Inventors: Douglas A. Treco, Michael W. Heartlein, Richard F Selden
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Patent number: 6225525Abstract: A transgenic mouse with alterations in an abc1 gene is prepared by introduction of an altered abc1 gene into a host animal. The resulting transgenic mice do not produce functional ABC1 protein molecules. Cells and cell lines derived from these animals also contain the altered abc1 gene.Type: GrantFiled: October 13, 1999Date of Patent: May 1, 2001Assignee: Ortho-McNeil Pharmaceutical, Inc.Inventors: Wai-Ping Leung, Trudy Christiansen-Weber, Joseph R. Voland, Per A. Peterson
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Patent number: 6218187Abstract: The invention provides a novel retroviral packaging system, in which retroviral packaging plasmids and packagable vector transcripts are produced from high expression plasmids after stable or transient transfection in mammalian cells. High titers of recombinant retrovirus are produced in these transfected mammalian cells and can then transduce a mammalian target cell by cocultivation or supernatant infection. The methods of the invention include the use of the novel retroviral packaging plasmids and vectors to transduce primary human cells, including T cells and human hematopoietic stem cells, with foreign genes by cocultivation or supernatant infection at high efficiencies. The invention is useful for the rapid production of high titer viral supernatants, and to transduce with high efficiency cells that are refractory to transduction by conventional means.Type: GrantFiled: March 11, 1999Date of Patent: April 17, 2001Assignee: Cell Genesys, Inc.Inventors: Mitchell H. Finer, Thomas J. Dull, Krisztina M. Zsebo, Keegan Cooke, Deborah A. Farson
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Patent number: 6214622Abstract: The present invention relates to a method of gene or DNA targeting in cells of vertebrate, particularly mammalian, origin. That is, it relates to a method of introducing DNA into primary or secondary cells of vertebrate origin through homologous recombination or targeting of the DNA, which is introduced into genomic DNA of the primary or secondary cells at a preselected site. The present invention further relates to primary or secondary cells, referred to as homologously recombinant (HR) primary or secondary cells, produced by the present method and to uses of the homologously recombinant primary or secondary cells. The present invention also relates to a method of turning on a gene present in primary cells, secondary cells or immortalized cells of vertebrate origin, which is normally not expressed in the cells or is not expressed at significant levels in the cells.Type: GrantFiled: May 18, 1995Date of Patent: April 10, 2001Assignee: Transkaryotic Therapies, Inc.Inventors: Douglas Treco, Michael W. Heartlein, Richard F Selden
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Patent number: 6210922Abstract: The present invention relates to a method of serum-free production of recombinant proteins and adenoviral vectors; cell lines viable in a serum-free medium and method of obtaining same.Type: GrantFiled: November 30, 1998Date of Patent: April 3, 2001Assignee: National Research Council of CanadaInventors: M. T. Johanne Côté, Amine A. Kamen, Bernard Massie
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Patent number: 6211351Abstract: The invention is based on the discovery that recombinagenic oligonucleobases are active in prokaryotic cells that contain a strand transfer activity (RecA) and mismatch repair activity (MutS). Using this system a type of Duplex Mutational Vector termed a Heteroduplex Mutational Vector, was shown to be more active in prokaryotic cells than the types of mutational vectors heretofore tested. Further improvements in activity were obtained by replacing the tetrathymidine linker by a nuclease resistant oligonucleotide, such as tetra-2′-O-methyl-uridine, to link the two strands of the recombinagenic oligonucleobase and removing the DNA-containing intervening segment. The claims concern Duplex Mutational Vectors that contain the above improvements. In an alternative embodiment the claims concern the use of Duplex Mutational Vectors in prokaryotic cells.Type: GrantFiled: October 28, 1999Date of Patent: April 3, 2001Assignee: ValiGen (US), Inc.Inventors: Ramesh Kumar, Richard A. Metz
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Patent number: 6207457Abstract: A novel targeted nucleotide sequence delivery and integration system is provided. The integration system employs nucleic acid constructs having a targeting sequence capable of binding AAV Rep and a heterologous nucleotide sequence arranged relative to the targeting sequence such that the targeting sequence is capable of directing the integration of the heterologous sequence into a target site in a recipient genome. The system further employs Rep expression products which provide integration functions effective to mediate the site-specific integration of the targeting sequence and the heterologous sequence into the recipient genome. Methods are described, whereby the nucleotide sequence integration system can deliver and efficiently integrate large nucleotide sequences into target sites in recipient cell genomes. Therapeutic methods are also provided, wherein the integration systems are used to insert various therapeutically relevant nucleotide sequences into selected cells from a subject.Type: GrantFiled: September 8, 1995Date of Patent: March 27, 2001Assignee: Avigen, Inc.Inventors: Georges Natsoulis, Richard T. Surosky
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Patent number: 6204062Abstract: The invention provides in one embodiment a composition which is a linear DNA molecule having a desired replacement sequence, and second and third sequences substantially homologous to non-identical portions of the gene and having proximal and distal ends, the proximal ends flanking the desired replacement sequence and the distal ends having a terminating nucleotide analog at each end of the molecule. Another embodiment of the invention provides methods, by blocking the 3′ ends of transforming DNA with 2′3′ dideoxynucleotides, to reduce the frequency of end-mediated DNA insertion. These methods introduce only one copy of the selectable gene at the target locus to achieve a precise gene disruption, reducing or eliminating undesirable and multiple insertions that occur both non-homologously and at the targeted locus.Type: GrantFiled: August 10, 1999Date of Patent: March 20, 2001Assignee: Trustees of Amherst CollegeInventor: David I. Ratner
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Patent number: 6204061Abstract: Positive-negative selector (PNS) vectors are provided for modifying a target DNA sequence contained in the genome of a target cell capable of homologous recombination. The vector comprises a first DNA sequence which contains at least one sequence portion which is substantially homologous to a portion of a first region of a target DNA sequence. The vector also includes a second DNA sequence containing at least one sequence portion which is substantially homologous to another portion of a second region of a target DNA sequence. A third DNA sequence is positioned between the first and second DNA sequences and encodes a positive selection marker which when expressed is functional in the target cell in which the vector is used. A fourth DNA sequence encoding a negative selection marker, also functional in the target cell, is positioned 5′ to the first or 3′ to the second DNA sequence and is substantially incapable of homologous recombination with the target DNA sequence.Type: GrantFiled: January 9, 1997Date of Patent: March 20, 2001Assignee: University of Utah Research FoundationInventors: Mario R. Capecchi, Kirk R. Thomas
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Patent number: 6204432Abstract: A transgenic mouse with alterations in a PA28&bgr; gene is prepared by introduction of an altered PA28&bgr; gene into a host mouse. The resulting transgenic animals do not produce functional PA28 molecules. Cells and cell lines derived from these animals also contain the altered PA28&bgr; gene.Type: GrantFiled: August 31, 1999Date of Patent: March 20, 2001Assignee: Ortho-McNeil Pharmaceutical, Inc.Inventors: Wai-Ping Leung, Young Yang, Tobias Preckel, Per Peterson
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Patent number: 6200812Abstract: The invention relates to methods for targeting an exogenous polynucleotide or exogenous complementary polynucleotide pair to a predetermined endogenous DNA target sequence in a target cell by homologous pairing, particularly for altering an endogenous DNA sequence, such as a chromosomal DNA sequence, typically by targeted homologous recombination. In certain embodiments, the invention relates to methods for targeting an exogenous polynucleotide having a linked chemical substituent to a predetermined endogenous DNA sequence in a metabolically active target cell, generating a DNA sequence-specific targeting of one or more chemical substituents in an intact nucleus of a metabolically active target cell, generally for purposes of altering a predetermined endogenous DNA sequence in the cell.Type: GrantFiled: April 8, 1999Date of Patent: March 13, 2001Assignee: SRI InternationalInventors: Sushma Pati, David A. Zarling
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Patent number: 6172278Abstract: Transgenic cells, transgenic mice having an Ikaros transgene and methods for the use thereof.Type: GrantFiled: July 29, 1994Date of Patent: January 9, 2001Assignee: The General Hospital CorporationInventor: Katia Georgopoulos
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Patent number: 6159736Abstract: A method for making insertional mutations at random or quasi-random locations in the chromosomal or extra-chromosomal nucleic acid of a target cell includes the step of combining, in the target cell, cellular nucleic acid with a synaptic complex that comprises (a) a Tn5 transposase protein and (b) a polynucleotide that comprises a pair of nucleotide sequences adapted for operably interacting with Tn5 transposase and a transposable nucleotide sequence therebetween, under conditions that mediate transpositions into the cellular DNA. In the method, the synaptic complex is formed in vitro under conditions that disfavor or prevent the synaptic complexes from undergoing productive transposition.Type: GrantFiled: September 23, 1998Date of Patent: December 12, 2000Assignee: Wisconsin Alumni Research FoundationInventors: William S. Reznikoff, Igor Y. Goryshin
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Patent number: 6159680Abstract: A targeted disruption of the NF1 gene in mice has been used to demonstrate that both neural crest- and placode-derived sensory neurons isolated from NF1(-/-) embryos develop, extend neurites, and survive in the absence of neurotrophins, whereas their wild-type counterparts die rapidly unless NGF or BDNF is added to the culture medium. Moreover, NF1 mutant sympathetic neurons survive for extended periods and acquire mature morphology in the presence of NGF-blocking antibodies. The discovery is useful in screening candidate substances for inhibition of neurofibromin action and in therapy for neurodegeneration due to disease or trauma.Type: GrantFiled: July 7, 1995Date of Patent: December 12, 2000Assignee: Board of Regents, The University of Texas SystemInventors: Luis F. Parada, Kristine S. Vogel
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Patent number: 6156574Abstract: A method of efficiently sequencing multiple exons from complex genomic DNAs is disclosed. The methodology includes the use of bacterial and bacteriophage-derived artificial chromosomes (BBPACs) in novel gene trapping protocols. Targeted gene trapping by homologous recombination, and random gene trapping with the use of a transposon system are exemplified. Included in the invention are methods of preparing a gene map from BBPAC contigs, the resulting gene maps, methods of constructing a cDNA library from BBPAC contigs, and the resulting cDNA libraries.Type: GrantFiled: June 22, 1998Date of Patent: December 5, 2000Assignee: The Rockefeller UniversityInventors: Nathaniel Heintz, Weining Jiang, Xiangdong W. Yang
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Patent number: 6156303Abstract: The present invention provides isolated adenovirus-associated viruses ("AAV"), including AAV isolates designated AAV3B and AAV6. The invention also provides nucleic acid molecules of AAV3B (SEQ ID NO: 1) or AAV6 (SEQ ID NO: 2), including DNA or RNA, and provides substantially purified polypeptides encoded by AAV3B or AAV6, as well as antibodies specific for such polypeptides. The invention also provides infectious AAV3B and AAV6 clones; AAV viral vectors, which can be hybrid AAV viral vectors; AAV vector plasmids; and AAV helper plasmids; each comprising at least a portion of an AAV3B (SEQ ID NO: 1) or AAV6 (SEQ ID NO: 2) nucleic acid molecule. The invention further provides host cells containing at least a portion of an AAV3B or AAV6 nucleic acid molecule and progeny cells derived therefrom, and provides non-human transgenic mammals having an AAV vector genome stably integrated in a chromosome.Type: GrantFiled: June 11, 1997Date of Patent: December 5, 2000Assignee: University of WashingtonInventors: David W. Russell, Elizabeth A. Rutledge
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Patent number: 6150169Abstract: This invention relates to DNA constructs for inserting heterologous gene sequences into a host genome so as to obtain expression of the heterologous gene, to methods of inserting heterologous gene sequences into a host genome, and to organisms carrying modified host genomes. Specifically, the DNA constructs of this invention contain an expression unit of an internal ribosome binding site (IRES) coupled to a heterologous gene sequence. This expression unit is flanked at the 5' and 3' ends by DNA sequences that enable homologous recombination or integration of the construct with the DNA of a targeted host to obtain expression of the heterologous gene in the host.Type: GrantFiled: January 25, 1996Date of Patent: November 21, 2000Assignee: The University of EdinburghInventors: Austin Gerard Smith, Peter Scott Mountford, Richard Frank Lathe
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Patent number: 6143566Abstract: A simple method for modifying genes in a recombination deficient host cell is disclosed. Such modifications include generating insertion, deletions, substitutions, and/or point mutations at any chosen site in the independent origin based cloning vector. The modified gene can be contained in an independent origin based cloning vector that is used to introduce a modified heterologous gene into a cell. Such a modified vector may be used in the production of a germline transmitted transgenic animal, or in gene targeting protocols in eukaryotic cells.Type: GrantFiled: June 23, 1997Date of Patent: November 7, 2000Assignee: The Rockfeller UniversityInventors: Nathaniel Heintz, Peter Model, Xiangdong W. Yang
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Patent number: 6139835Abstract: Homologous recombination is employed to inactivate genes, particularly genes associated with MHC antigens. Particularly, the .beta..sub.2- microglobulin gene is inactivated for reducing or eliminating the expression of functional Class I MHC antigens. The resulting cells may be used as allogeneic donor cells. Methods for homologous recombination in non-transformed mammalian somatic cells are also described.Type: GrantFiled: May 18, 1995Date of Patent: October 31, 2000Assignee: Cell Genesys, Inc.Inventors: Raju Kucherlapati, Beverly H. Koller, Oliver Smithies, Robert B. Dubridge, Gary Greenburg, Daniel J. Capon, Steven R. Williams, Mariona Lourdes Arbones De Rafael
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Patent number: 6130364Abstract: A method to produce a cell expressing an antibody from a genomic sequence of the cell comprising a modified immunoglobulin locus using Cre-mediated site-specific recombination is disclosed. The method involves first transfecting an antibody-producing cell with a homology-targeting vector comprising a lox site and a targeting sequence homologous to a first DNA sequence adjacent to the region of the immunoglobulin loci of the genomic sequence which is to be converted to a modified region, so the first lox site is inserted into the genomic sequence via site-specific homologous recombination. Then the cell is transfected with a lox-targeting vector comprising a second lox site suitable for Cre-mediated recombination with the integrated lox site and a modifying sequence to convert the region of the immunoglobulin loci to the modified region.Type: GrantFiled: March 29, 1995Date of Patent: October 10, 2000Assignees: Abgenix, Inc., Japan Tobacco Inc.Inventors: Aya Jakobovits, Krisztina M. Zsebo
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Patent number: 6130090Abstract: A method of efficiently sequencing multiple exons from complex genomic DNAs is disclosed. The methodology includes the use of bacterial and bacteriophage-derived artificial chromosomes (BBPACs) in novel gene trapping protocols. Targeted gene trapping by homologous recombination, and random gene trapping with the use of a transposon system are exemplified. Included in the invention are methods of preparing a gene map from BBPAC contigs, the resulting gene maps, methods of constructing a cDNA library from BBPAC contigs, and the resulting cDNA libraries.Type: GrantFiled: January 14, 1998Date of Patent: October 10, 2000Assignee: The Rockefeller UniversityInventors: Nathaniel Heintz, Weining Jiang, Xiangdong W. Yang
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Patent number: 6127598Abstract: The present invention features mouse models for Nkx-2.2 gene function and for Nkx-6.1 gene function, wherein the transgenic mouse is characterized by having a defect in Nkx-2.2 gene function or a defect in Nkx-6.1 gene function (where, because Nkx-2.2 acts upstream of Nkx-6.1, a defect in Nkx-2.2 gene function affects Nkx-6.1 gene function) and by having a decreased number of insulin-producing cells relative to a normal mouse. Where the transgenic mouse contains a defect in Nkx-2.2 gene function, the mouse is further characterized by a decreased number of serotonin-producing cells relative to a normal mouse. The transgenic mice may be either homozygous or heterozygous for the Nkx-2.2 or Nkx-6.1 defect.Type: GrantFiled: July 25, 1997Date of Patent: October 3, 2000Assignee: The Regents of the University of CaliforniaInventors: Michael S. German, John L.R. Rubenstein, Lori Sussel, Maike Sander, Dennis J. Hartigan-O'Connor, Roger A. Pedersen, Juanito J. Meneses
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Patent number: 6100445Abstract: A transgenic mouse having somatic and germ cells in which at least one allele of an endogenous interleukin-1.beta. converting enzyme (ICE) gene is functionally disrupted is provided. The mouse may be heterozygous or, more preferably, homozygous for the ICE gene disruption. In homozygous mice, secretion of mature interleukin-1.beta. and interleukin-1.alpha. is substantially reduced relative to non-mutant mice. The mice of the invention can be used as positive controls to evaluate the efficacy of ICE inhibitors and to identify disease conditions that can be treated with ICE inhibitors. A transgenic mouse having functionally disrupted endogenous ICE genes but which has been reconstituted with a human ICE gene is also provided. This mouse can be used to identify agents that inhibit human ICE in vivo.Type: GrantFiled: October 20, 1997Date of Patent: August 8, 2000Assignee: BASF AktiengesellschaftInventors: Tara Seshadri, Ping Li, Hamish Allen, Subhashis Banerjee, Michael Paskind
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Patent number: 6093873Abstract: The present invention is directed to mice which are genetically altered to be deficient in the normal expression of RXR.gamma., to mice heterozygous for such deficiency, and to cell lines, preferably pluripotent or totipotent cell lines, which are heterozygous or homozygous for such deficiency. The invention further provides mice and cell lines which, in addition to being deficient in RXR.gamma., are genetically altered to be deficient in the expression of RXR.alpha. and/or RXR.beta.. The present invention further provides the use of any of the above mice and cell lines in situations where the absence of RXR.gamma., or the normal expression thereof, is desirable.Type: GrantFiled: August 19, 1997Date of Patent: July 25, 2000Assignees: Institut National de la Sante et de la Recherche Medicale, Centre National de la Recherche Scientifique, Universite Louis Pasteur, Bristol-Myers Squibb CompanyInventors: Pierre Chambon, Philippe Kastner
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Patent number: 6087555Abstract: Disclosed is a mouse in which expression of the gene encoding Osteoprotegerin is suppressed. Also disclosed is a nucleic acid construct useful in preparing such a mouse, and a cell line containing such construct.Type: GrantFiled: October 15, 1997Date of Patent: July 11, 2000Assignee: Amgen Inc.Inventors: Colin Dunstan, Scott Simonet, Ildiko Sarosi
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Patent number: 6080911Abstract: The present invention provides mouse models of growth hormone insensitivity including Laron syndrome. In particular, the present invention provides transgenic mice incapable of expressing functional growth hormone receptor including mice which further cannot express functional growth hormone binding protein. The invention further provides methods for testing the usefulness of chemical compounds in the treatment of growth hormone insensitivity and diabetic end-organ disease.Type: GrantFiled: April 15, 1997Date of Patent: June 27, 2000Assignee: Ohio UniversityInventors: John J. Kopchick, Yihua Zhou
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Patent number: 6080576Abstract: A novel 3' gene trap cassette is described that does not encode a marker conferring antibiotic resistance and can be used to efficiently trap and identify cellular genes. Vectors incorporating the presently 3' gene trap cassette find particular application in gene discovery, the production of transgenic cells and animals, and gene activation.Type: GrantFiled: April 8, 1998Date of Patent: June 27, 2000Assignee: Lexicon Genetics IncorporatedInventors: Brian Zambrowicz, Glenn Friedrich, Arthur T. Sands