Involving Site-specific Recombination (e.g., Cre-lox, Etc.) Patents (Class 435/462)
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Publication number: 20150064789Abstract: Disclosed herein are compositions for linking DNA binding domains and cleavage domains (or cleavage half-domains) to form non-naturally occurring nucleases. Also described are methods of making and using compositions comprising these linkers.Type: ApplicationFiled: August 28, 2014Publication date: March 5, 2015Inventors: David Paschon, Lei Zhang
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Publication number: 20150056705Abstract: Methods and compositions for genetic alteration of cells are provided.Type: ApplicationFiled: May 15, 2014Publication date: February 26, 2015Inventors: Anthony Conway, Gregory J. Cost, Russell Dekelver, Edward J. Rebar, Andreas Reik, Fyodor Urnov, Jianbin Wang, H. Steve Zhang
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Publication number: 20150059010Abstract: Compositions and methods are provided for genome modification of a target sequence in the genome of a cell. The methods and compositions employ a guide polynucleotide/Cas endonuclease system to provide an effective system for modifying or altering target sites within the genome of a cell or organism. Once a genomic target site is identified, a variety of methods can be employed to further modify the target sites such that they contain a variety of polynucleotides of interest. Compositions and methods are also provided for editing a nucleotide sequence in the genome of a cell. Breeding methods and methods for selecting plants utilizing a two component RNA polynucleotide and Cas endonuclease system are also disclosed.Type: ApplicationFiled: August 20, 2014Publication date: February 26, 2015Inventors: Andrew Mark Cigan, Phillip A. Patten, Joshua K. Young
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Publication number: 20150050699Abstract: Isolation or in vitro assembly of the Cas9-crRNA complex of the Streptococcus thermophilus CRISPR3/Cas system and use for cleavage of DNA bearing a nucleotide sequence complementary to the crRNA and a proto-spacer adjacent motif. Methods for site-specific modification of a target DNA molecule in vitro or in vivo using an RNA-guided DNA endonuclease comprising RNA sequences and at least one of an RuvC active site motif and an HNH active site motif; for conversion of Cas9 polypeptide into a nickase cleaving one strand of double-stranded DNA by inactivating one of the active sites (RuvC or HNH) in the polypeptide by at least one point mutation; for assembly of active polypeptide-polyribonucleotides complex in vivo or in vitro; and for re-programming a Cas9-crRNA complex specificity in vitro and using a cassette containing a single repeat-spacer-repeat unit.Type: ApplicationFiled: March 20, 2013Publication date: February 19, 2015Inventors: Virginijus Siksnys, Giedrius Gasiunas, Tautvydad Karvelis, Arvydas Lubys, Lolita Zaliauskiene, Monika Glemzaite, Anja Smith
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Publication number: 20150050739Abstract: The present invention relates to a method for carrying out recombination at a target locus.Type: ApplicationFiled: March 12, 2013Publication date: February 19, 2015Inventors: Noel Nicolaas Maria Elisabeth Van Peij, Martina Beishuizen, Yvonne Johannes Odilia Arendsen
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Publication number: 20150044772Abstract: An inactive CRISPR/Cas system-based fusion protein and its applications in gene editing are disclosed. More particularly, chimeric fusion proteins including an inCas fused to a DNA modifying enzyme and methods of using the chimeric fusion proteins in gene editing are disclosed. The methods can be used to induce double-strand breaks and single-strand nicks in target DNAs, to generate gene disruptions, deletions, point mutations, gene replacements, insertions, inversions and other modifications of a genomic DNA within cells and organisms.Type: ApplicationFiled: August 8, 2014Publication date: February 12, 2015Inventor: Guojun Zhao
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Publication number: 20150037892Abstract: The present invention relates to a method for carrying out recombination at a target locus.Type: ApplicationFiled: March 12, 2013Publication date: February 5, 2015Inventors: Nathalie Wiessenhaan, Catharina Petronella Antonia Maria Kolen, Bernard Meijrink, Viktor Marius Boer, Johannes Andries Roubos, Yvonne Johannes Odilia Arendsen
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Patent number: 8945884Abstract: The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different.Type: GrantFiled: May 13, 2013Date of Patent: February 3, 2015Assignee: Life Technologies CorporationInventors: Jonathan Chesnut, John Carrino, Louis Leong, Knut Madden, Martin Gleeson, James Fan, Michael Brasch, David Cheo, James Hartley, Devon Byrd, Gary Temple
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Publication number: 20150031132Abstract: Methods of genome engineering in cells using a TALEN lacking repeat sequences or Cas9 is provided.Type: ApplicationFiled: June 30, 2014Publication date: January 29, 2015Inventors: George M. CHURCH, Luhan YANG, Marc Guell CARGOL, Joyce Lichi YANG
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Publication number: 20150031090Abstract: Disclosed herein are compositions and methods for generating chromosomal translocations and targeted deletions of specific lengths and at specific locations the genome of cell.Type: ApplicationFiled: September 30, 2014Publication date: January 29, 2015Inventors: Fyodor Urnov, Jianbin Wang
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Publication number: 20150031133Abstract: Methods of genome engineering in cells using a TALEN lacking repeat sequences or Cas9 is provided.Type: ApplicationFiled: June 30, 2014Publication date: January 29, 2015Inventors: George M. CHURCH, Luhan YANG, Marc Guell CARGOL, Joyce Lichi YANG
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Publication number: 20150023943Abstract: Purified Ref polypeptides with increased nuclease site-specific targeting activity, recombinant nucleic acids and cells for expression of such Ref polypeptides, and methods for using the Ref polypeptides in combination with RecA protein and variants thereof to effect targeted nuclease cleavage of a DNA duplex are disclosed.Type: ApplicationFiled: July 21, 2014Publication date: January 22, 2015Inventors: Michael Matthew Cox, Angela Jo Gruber, Tayla Maria Olsen
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Publication number: 20150024500Abstract: The present invention provides a method and compositions utilizing the CRISPR system to disrupt a target gene in eukaryotic cells to produce double allele knock outs. The method finds use in producing afucosylated antibodies with enhanced ADCC activity.Type: ApplicationFiled: July 19, 2014Publication date: January 22, 2015Applicant: LARIX BIOSCIENCE, LLCInventors: Bo Yu, James Larrick
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Publication number: 20150024499Abstract: A clustered regularly interspaced short palindromic repeat (CRISPR)-associated complex for adaptive antiviral defence (Cascade); the Cascade protein complex comprising at least CRISPR-associated protein subunits Cas7, Cas5 and Cas6 which includes at least one subunit with an additional amino acid sequence possessing nucleic acid or chromatin modifying, visualising, transcription activating or transcription repressing activity. The Cascade complex with additional activity is combined with an RNA molecule to produce a ribonucleoprotein complex. The RNA molecule is selected to have substantial complementarity to a target sequence. Targeted ribonucleoproteins can be used as genetic engineering tools for precise cutting of nucleic acids in homologous recombination, non-homologous end joining, gene modification, gene integration, mutation repair or for their visualisation, transcriptional activation or repression.Type: ApplicationFiled: July 8, 2014Publication date: January 22, 2015Inventors: Stan Johan Jozef Brouns, John Van Der Oost
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Publication number: 20150017728Abstract: The present invention provides compositions and methods for targeted cleavage of cellular chromatin in a region of interest and/or homologous recombination at a predetermined site in cells. Compositions include fusion polypeptides comprising a TAL effector binding or a zinc finger domain and an I-TevI homing endonuclease cleavage domain as well as nucleic acid sequence encoding the same. The use of the I-TevI domain allows for monomer endonuclease sequences to achieve cleavage of cellular chromatin and represents an advantage over prior endonucleases which require self-dimerization, and two nucleases with appropriate spacers.Type: ApplicationFiled: September 19, 2012Publication date: January 15, 2015Inventors: Bing Yang, Ting Li, Sheng Huang
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Publication number: 20140359796Abstract: A genetically modified livestock animal, and methods of making and using the same, the animal comprising a genetic modification to disrupt a target gene selectively involved in gametogenesis, wherein the disruption of the target gene prevents formation of functional gametes of the animal. Animals that create progeny with donor genetics, and methods of making and using the same. Cells, and methods of making and using the cells, with a genetic modification to disrupt a target gene selectively involved in gametogenesis.Type: ApplicationFiled: April 28, 2014Publication date: December 4, 2014Inventors: Scott C. Fahrenkrug, Daniel F. Carlson
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Publication number: 20140359798Abstract: Genetically engineered mice having a corrected Cbr1rd8 mutation and methods for correcting the Cbr1rd8 mutation to produce the genetically engineered mice are provided according to aspects of the present invention. Kits, expression vectors and fusion proteins according to aspects of the invention are provided for use to produce the genetically engineered mice characterized by a corrected Cbr1rd8 mutation.Type: ApplicationFiled: June 3, 2014Publication date: December 4, 2014Applicant: THE JACKSON LABORATORYInventors: Michael V. Wiles, Benjamin E. Low
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Publication number: 20140356960Abstract: The present invention relates to a nucleic acid containing at least one homing endonuclease site (HE) and at least one restriction enzyme site (X) wherein the HE and X sites are selected such that HE and X result in compatible cohesive ends when cut by the homing endonuclease and restriction enzyme, respectively, and the ligation product of HE and X cohesive ends can neither be cleaved by the homing endonuclease nor by the restriction enzyme. Further subject-matter of the present invention relates to a vector comprising the nucleic acid of the present invention, host cells containing the nucleic acid and/or the vector; a kit for cloning and/or expression of multiprotein complexes making use of the vector and the host cells, a method for producing a vector containing multiple expression cassettes, and a method for producing multiprotein complexes.Type: ApplicationFiled: April 25, 2014Publication date: December 4, 2014Inventor: Imre Berger
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Publication number: 20140351965Abstract: Compositions and methods for swine LDL-R gene knockouts.Type: ApplicationFiled: August 15, 2014Publication date: November 27, 2014Inventors: Scott C Fahrenkrug, Daniel F Carlson
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Publication number: 20140349405Abstract: Disclosed are components and methods for RNA-directed DNA cleavage and gene editing. The components include and the methods utilize a Cas9 protein from Neisseria and one or more RNA molecules in order to direct the Cas9 protein to bind to and optionally cleave or nick a target sequence.Type: ApplicationFiled: May 22, 2014Publication date: November 27, 2014Applicants: WISCONSIN ALUMNI RESEARCH FOUNDATION, NORTHWESTERN UNIVERSITYInventors: Erik J. Sontheimer, Yan Zhang, Alfonso Mondragon, Rakhi Rajan, James Thomson, Zhonggang Hou
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Publication number: 20140342458Abstract: A method of altering a eukaryotic cell is provided including transfecting the eukaryotic cell with a nucleic acid encoding RNA complementary to genomic DNA of the eukaryotic cell, transfecting the eukaryotic cell with a nucleic acid encoding an enzyme that interacts with the RNA and cleaves the genomic DNA in a site specific manner, wherein the cell expresses the RNA and the enzyme, the RNA binds to complementary genomic DNA and the enzyme cleaves the genomic DNA in a site specific manner.Type: ApplicationFiled: June 30, 2014Publication date: November 20, 2014Inventors: Prashant G. MALI, George M. CHURCH, Luhan Yang
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Publication number: 20140335063Abstract: Disclosed herein are methods and compositions for delivery of engineered nucleases and donor molecules into the genome of a cell.Type: ApplicationFiled: May 6, 2014Publication date: November 13, 2014Inventors: Paula M. Cannon, Colin Michael Exline, Michael C. Holmes
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Patent number: 8877435Abstract: Methods and means are provided for producing chimeric nucleic acid constructs capable of producing dsRNA for silencing target nucleic acid sequences of interest using recombinational cloning.Type: GrantFiled: September 21, 2010Date of Patent: November 4, 2014Assignee: Commonwealth Scientific and Industrial Research OrganisationInventors: Christopher A. Helliwell, Susan V. Wesley, Peter M. Waterhouse
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Patent number: 8871516Abstract: The present invention is directed to a method for preparing an expression vector encoding a tailored recombinase, wherein said tailored recombinase recombines asymmetric target sites within the LTR of proviral DNA of a retrovirus inserted into the genome of a host cell and is useful as means for excising the provirus from the genome of the host cell. The present invention further relates to an in vitro-method of optimising the treatment of a retroviral infection of a subject and to the use of tailored recombinases for the preparation of pharmaceutical compositions for reducing the viral load in a subjected infected by a retrovirus.Type: GrantFiled: January 3, 2008Date of Patent: October 28, 2014Assignees: Technische Universität Dresden, Max-Planck-Gesellschaft zur Förderung der Wissenschaften E.V., Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie an der Universität HamburgInventors: Joachim Hauber, Frank Buchholz, Ilona Hauber, Francis A Stewart, Indrani Sarkar
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Patent number: 8871905Abstract: Disclosed herein are methods and compositions for modulating activity of CXCR4 genes, for example using zinc finger transcription factors (ZF-TFs) or zinc finger nucleases (ZFNs) comprising a zinc finger protein and a cleavage domain or cleavage half-domain. Polynucleotides encoding ZF-TFs or ZFNs, vectors comprising polynucleotides encoding ZF-TFs or ZFNs and cells comprising polynucleotides encoding ZF-TFs or ZFNs and/or cells comprising ZF-TF or ZFNs are also provided.Type: GrantFiled: March 18, 2010Date of Patent: October 28, 2014Assignee: Sangamo BioSciences, Inc.Inventors: Michael C. Holmes, Jeffrey C. Miller, Jianbin Wang
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Publication number: 20140315985Abstract: This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof.Type: ApplicationFiled: March 12, 2014Publication date: October 23, 2014Applicant: Caribou Biosciences, Inc.Inventors: Andrew Paul May, Rachel E. Haurwitz, Jennifer A. Doudna, James M. Berger, Matthew Merrill Carter, Paul Donohoue
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Publication number: 20140309487Abstract: Compositions and methods are provided for modifying a rat genomic locus of interest using a large targeting vector (LTVEC) comprising various endogenous or exogenous nucleic acid sequences as described herein. Compositions and methods for generating a genetically modified rat comprising one or more targeted genetic modifications in their germline are also provided. Compositions and methods are provided which comprise a genetically modified rat or rat cell comprising a targeted genetic modification in the rat interleukin-2 receptor gamma locus, the rat ApoE locus, the rat Rag2 locus, the rat Rag1 locus and/or the rat Rag2/Rag1 locus. The various methods and compositions provided herein allows for these modified loci to be transmitted through the germline.Type: ApplicationFiled: June 25, 2014Publication date: October 16, 2014Inventors: Jeffrey D. Lee, Alexander O. Mujica, Wojtek Auerbach, Ka-Man Venus Lai, David M. Valenzuela, George D. Yancopoulos
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Publication number: 20140310828Abstract: Compositions and methods are provided for modifying a rat genomic locus of interest using a large targeting vector (LTVEC) comprising various endogenous or exogenous nucleic acid sequences as described herein. Compositions and methods for generating a genetically modified rat comprising one or more targeted genetic modifications in their germline are also provided. Compositions and methods are provided which comprise a genetically modified rat or rat cell comprising a targeted genetic modification in the rat interleukin-2 receptor gamma locus, the rat ApoE locus, the rat Rag2 locus, the rat Rag1 locus and/or the rat Rag2/Rag1 locus. The various methods and compositions provided herein allows for these modified loci to be transmitted through the germline.Type: ApplicationFiled: April 16, 2014Publication date: October 16, 2014Inventors: Jeffrey D. Lee, Alexander O. Mujica, Wojtek Auerbach, Ka-Man Venus Lai, David M. Valenzuela, George D. Yancopoulos
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Publication number: 20140304847Abstract: The present invention relates to a method for modifying a target sequence in the genome of a mammalian cell, the method comprising the step of introducing into a mammalian cell: a. one or more compounds that introduce double-strand breaks in said target sequence; b. one or more DNA molecules comprising a donor DNA sequence to be incorporated by homologous recombination into the genomic DNA of said mammalian cell within said target sequence, wherein said donor DNA sequence is flanked upstream by a first flanking element and downstream by a second flanking element, wherein said first and second flanking element are different and wherein each of said first and second flanking sequence are homologous to a continuous DNA sequence on either side of the double-strand break introduced by said one or more compounds of a. within said target sequence in the genome of said mammalian cell; and c.Type: ApplicationFiled: June 6, 2012Publication date: October 9, 2014Inventors: Ralf Kühn, Wolfgang Wurst
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Publication number: 20140287513Abstract: The present invention is directed to nucleic acid and amino acid sequences of a novel piggyBac transposase enzymes created by modifying the transposase of Trichoplusia ni. The piggyBac transposases of the present invention are functionally active or hyperactive for excision and have decreased integration activity compared to wild type Trichoplusia ni piggyBac transposase enzyme. These transposases are ideal for use in methods of transforming cells and organisms. In particular embodiments, the present invention provides methods of transient integration and expression of transgenes.Type: ApplicationFiled: July 16, 2012Publication date: September 25, 2014Applicants: HOWARD HUGUES MEDICAL INSTITUTE, THE JOHNS HOPKINS UNIVERSITYInventor: Nancy L. Craig
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Publication number: 20140273234Abstract: The invention provides for engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are compositions and methods related to components of a CRISPR complex particularly comprising a Cas ortholog enzyme.Type: ApplicationFiled: June 2, 2014Publication date: September 18, 2014Applicants: The Board Institute, Inc., Massachusetts Institute of TechnologyInventors: Feng ZHANG, Fei RAN
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Publication number: 20140273232Abstract: The invention provides for systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for selecting specific cells by introducing precise mutations utilizing the CRISPR-Cas system.Type: ApplicationFiled: May 29, 2014Publication date: September 18, 2014Applicants: THE BROAD INSTITUTE, INC., MASSACHUSETTS INSTITUTE OF TECHNOLOGY, PRESIDENT AND FELLOWS OF HARVARD COLLEGEInventors: Feng Zhang, Le Cong, Patrick Hsu, Fei Ran
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Publication number: 20140273233Abstract: The present invention provides methods for modifying chromosomal sequences. In particular, methods are provided for using RNA-guided endonucleases or modified RNA-guided endonucleases to modify targeted chromosomal sequences.Type: ApplicationFiled: May 30, 2014Publication date: September 18, 2014Applicant: SIGMA-ALDRICH CO., LLCInventors: Fuqiang Chen, Gregory Davis
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Publication number: 20140273231Abstract: The invention provides for systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for selecting specific cells by introducing precise mutations utilizing the CRISPR/Cas system.Type: ApplicationFiled: April 23, 2014Publication date: September 18, 2014Applicants: THE BROAD INSTITUTE, INC., MASSACHUSETTS INSTITUTE OF TECHNOLOGYInventors: Feng ZHANG, Le CONG
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Publication number: 20140273230Abstract: The present invention provides methods for modifying chromosomal sequences. In particular, methods are provided for using RNA-guided endonucleases or modified RNA-guided endonucleases to modify targeted chromosomal sequences.Type: ApplicationFiled: March 14, 2014Publication date: September 18, 2014Applicant: SIGMA-ALDRICH CO., LLCInventors: Fuqiang Chen, Gregory Davis
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Publication number: 20140273037Abstract: The invention relates to engineered CRISPR/Cas9 systems for genomic modification in mammalian cells. The present specification describes the design and testing of a polynucleotide encoding the Streptococcus pyogenes (S. pyogenes) Cas9 protein, where the nucleotide sequence has been optimized for expression in mammalian cells. The specification also describes all-in-one systems for RNA-guided genome engineering in mammalian cells, including human cells.Type: ApplicationFiled: March 14, 2014Publication date: September 18, 2014Applicant: System Biosciences, LLCInventor: Fangting Wu
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Publication number: 20140283156Abstract: The present invention provides a trans-splicing ribozyme comprising i) a targeting nucleotide sequence that is complementary to a target nucleotide sequence within a mRNA that is expressed in a cell; contiguous with ii) a catalytic RNA sequence; contiguous with iii) a donor transcript, which donor transcript comprises at least a nucleotide sequence that encodes a trans-activator, wherein when the trans-splicing ribozyme is expressed in a cell, the catalytic RNA sequence cleaves the mRNA and ligates the donor transcript to the mRNA to generate a spliced mRNA which comprises the donor transcript, such that the donor transcript is translated as part of the spliced mRNA in the cell, as well as methods of using the trans-splicing ribozyme. The present invention also provides variants of Cre and other recombinases, as well as method of using the variants.Type: ApplicationFiled: March 13, 2014Publication date: September 18, 2014Applicant: COLD SPRING HARBOR LABORATORYInventors: Anthony M. Zador, Ian D. Peikon
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Publication number: 20140256798Abstract: The invention is directed to transcription activator-like effector nuclease (TALEN)-mediated DNA editing of disease-causing mutations in the context of the human genome and human cells to treat patients with compromised genetic disorders.Type: ApplicationFiled: February 28, 2014Publication date: September 11, 2014Inventors: Mark J. Osborn, Jakub Tolar, Bruce Blazar, Daniel F. Voytas
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Publication number: 20140248702Abstract: The invention provides for systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for selecting specific cells by introducing precise mutations utilizing the CRISPR/Cas system.Type: ApplicationFiled: April 22, 2014Publication date: September 4, 2014Applicants: THE BROAD INSTITUTE, INC., MASSACHUSETTS INSTITUTE OF TECHNOLOGYInventors: Feng ZHANG, Le CONG
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Publication number: 20140242702Abstract: Methods and compositions for increasing nuclease-mediated genomic modification using DNA repair inhibitors are provided.Type: ApplicationFiled: February 24, 2014Publication date: August 28, 2014Inventors: Fuqiang Chen, Qiaohua Kang, Thomas Wechsler
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Publication number: 20140227787Abstract: The invention provides for systems, methods, and compositions for altering expression of target gene sequences and related gene products. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for utilizing the CRISPR-Cas system.Type: ApplicationFiled: April 18, 2014Publication date: August 14, 2014Applicants: THE BROAD INSTITUTE, INC., MASSACHUSETTS INSTITUTE OF TECHNOLOGYInventor: Feng Zhang
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Patent number: 8790896Abstract: The present invention relates to a method for increasing the expression of a protein in cells, preferably in eukaryotic cells, by reducing the number of RNase L cleavage sites in the coding and/or non-coding region of the nucleic acid sequence of said protein. Furthermore, it relates to nucleic acid sequences exhibiting a reduced number of RNase L cleavage sites as well as to the proteins translated from such sequences.Type: GrantFiled: January 19, 2010Date of Patent: July 29, 2014Assignees: King Faisal Specialist Hospital & Research Centre, Terramark Markencreation GmbHInventor: Khalid S. Khabar
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Publication number: 20140205579Abstract: Nucleases and methods of using these nucleases for alteration of a CFTR gene and generation of cells and animal models.Type: ApplicationFiled: March 25, 2014Publication date: July 24, 2014Applicants: Board of Regents of the University of Texas System, Sangamo BioSciences, Inc.Inventors: Brian R. Davis, Dmitry M. Guschin, Michael C. Holmes, David Paschon, Phillip Tam
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Publication number: 20140208457Abstract: The present invention is concerned with nuclease fusion proteins and various uses thereof. Specifically, it relates to a polynucleotide encoding a polypeptide comprising (i) a first module comprising at least a first DNA binding domain derived from a homing endonuclease, (ii) a linker, and (iii) a second module comprising at least a second DNA binding domain and a cleavage domain derived from a restriction endonuclease, wherein said polypeptide functionally interacts only with DNA comprising a DNA recognition site for the first DNA binding domain and a DNA recognition site for the second DNA binding domain, and wherein said cleavage domain cleaves DNA within a specific DNA cleavage site upon binding of the polypeptide. Further contemplated are a vector and a non-human transgenic organism comprising said polynucleotide as well as a polypeptide encoded by the polynucleotide of the invention.Type: ApplicationFiled: June 8, 2012Publication date: July 24, 2014Applicant: BASF PLANT SCIENCE COMPANY GMBHInventors: Ines Fonfara, Wolfgang Wende, Alfred Pingoud
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Publication number: 20140193914Abstract: Methods and kits for lentiviral production are provided. By separating certain components of the vector including but not limited to promoter, reporter and selection marker preference components from the genetic content of interest, one can create libraries that allow for the efficient generation of custom selected lentiviral vectors.Type: ApplicationFiled: September 13, 2012Publication date: July 10, 2014Applicant: THERMO FISHER SCIENTIFIC BIOSCIENCES INC.Inventors: Jamie Cearley, Rusla DuBreuil
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Publication number: 20140186958Abstract: The invention provides for engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors with additional functional domains. Also provided are methods of directing CRISPR complex formation in prokaryotic and eukaryotic cells to ensure enhanced specificity for target recognition and avoidance of toxicity.Type: ApplicationFiled: December 12, 2013Publication date: July 3, 2014Inventors: Feng Zhang, Le Cong, Randall Jeffrey Platt, Neville Espi Sanjana, Fei Ran
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Publication number: 20140179005Abstract: Methods of inserting genes into defined locations in the chromosomal DNA of cultured mammalian cell lines which are subject to gene amplification are disclosed. In particular, sequences of interest (e.g., genes encoding biotherapeutic proteins) are inserted proximal to selectable genes in amplifiable loci, and the transformed cells are subjected to selection to induce co-amplification of the selectable gene and the sequence of interest. The invention also relates to meganucleases, vectors and engineered cell lines necessary for performing the methods, to cell lines resulting from the application of the methods, and use of the cell lines to produce protein products of interest.Type: ApplicationFiled: November 27, 2013Publication date: June 26, 2014Applicant: PRECISION BIOSCIENCES, INC.Inventors: Derek JANTZ, James Jefferson SMITH, Michael G. NICHOLSON
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Publication number: 20140179006Abstract: The invention provides for systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for selecting specific cells by introducing precise mutations utilizing the CRISPR/Cas system.Type: ApplicationFiled: February 18, 2014Publication date: June 26, 2014Applicants: Massachusetts Institute of Technology, The Broad Institute, Inc.Inventor: Feng ZHANG
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Publication number: 20140170753Abstract: The invention provides for systems, methods, and compositions for altering expression of target gene sequences and related gene products. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for utilizing the CRISPR-Cas system.Type: ApplicationFiled: February 18, 2014Publication date: June 19, 2014Applicants: MASSACHUSETTS INSTITUTE OF TECHNOLOGY, THE BROAD INSTITUTE INC.Inventor: Feng Zhang
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Publication number: 20140171494Abstract: There are provided methods and compositions for activating the expression of an exogenous gene by an exogenous integrase specifically in cells in which the exogenous integrase is expressed. The invention further relates to uses of the compositions in treatment of various conditions and disorders, as exemplified by selectively activating expression of a toxin only in target cell populations.Type: ApplicationFiled: August 1, 2012Publication date: June 19, 2014Applicant: RAMOT AT TEL-AVIV UNIVERSITY LTD.Inventors: Mikhail Kolot, Ezra Yagil, Natalia Malchin