Involving Site-specific Recombination (e.g., Cre-lox, Etc.) Patents (Class 435/462)
  • Patent number: 11060083
    Abstract: 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: Grant
    Filed: May 19, 2017
    Date of Patent: July 13, 2021
    Assignee: Larix Bioscience LLC
    Inventors: Bo Yu, James Larrick
  • Patent number: 11020430
    Abstract: This disclosure relates to the genetic modification of DNMT3A gene in immune cells. In certain embodiments, the modified immune cells may be used in adoptive T cells therapies to enhance immune responses against cancer or chronic infections. In certain embodiments, the disclosure relates to deleting, changing, or inserting nucleotides within the DNMT3A gene in immune cells, e.g., human CD8 T cells, such that the DNMT3A gene product does not function for methylation. In certain embodiments, modification of the DNMT3A gene provides an improvement in antigen-specific T cells functions and/or an enhancement of the longevity of the cells.
    Type: Grant
    Filed: November 4, 2016
    Date of Patent: June 1, 2021
    Assignees: Emory University, St. Jude Children's Research Hospital, Inc.
    Inventors: Rafi Ahmed, Benjamin Youngblood
  • Patent number: 11001829
    Abstract: The invention provides for systems, methods, and compositions for altering expression of target gene sequences and related gene products. Provided are structural information on the Cas protein of the CRISPR-Cas system, use of this information in generating modified components of the CRISPR complex, vectors and vector systems which encode one or more components or modified components of a CRISPR complex, as well as methods for the design and use of such vectors and components. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for utilizing the CRISPR-Cas system. In particular the present invention comprehends optimized functional CRISPR-Cas enzyme systems.
    Type: Grant
    Filed: March 24, 2017
    Date of Patent: May 11, 2021
    Assignees: The Broad Institute, Inc., Massachusetts Institute of Technology, President and Fellows of Harvard College
    Inventors: Feng Zhang, Silvana Konermann, Mark D. Brigham, Alexandra Trevino
  • Patent number: 10858639
    Abstract: Some aspects of this disclosure provide compositions, methods, and kits for improving the specificity of RNA-programmable endonucleases, such as Cas9. Also provided are variants of Cas9, e.g., Cas9 dimers and fusion proteins, engineered to have improved specificity for cleaving nucleic acid targets. Also provided are compositions, methods, and kits for site-specific nucleic acid modification using Cas9 fusion proteins (e.g., nuclease-inactivated Cas9 fused to a nuclease catalytic domain or a recombinase catalytic domain). Such Cas9 variants are useful in clinical and research settings involving site-specific modification of DNA, for example, genomic modifications.
    Type: Grant
    Filed: September 5, 2014
    Date of Patent: December 8, 2020
    Assignee: President and Fellows of Harvard College
    Inventors: David R. Liu, John Paul Guilinger, David B. Thompson
  • Patent number: 10787654
    Abstract: The present invention is directed to methods and compositions for genome editing and DNA targeting of proteins.
    Type: Grant
    Filed: January 23, 2015
    Date of Patent: September 29, 2020
    Assignee: NORTH CAROLINA STATE UNIVERSITY
    Inventors: Rodolphe Barrangou, Kurt M. Selle, Alexandra E. Briner
  • Patent number: 10745714
    Abstract: Disclosed herein are Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas) 9-based system related compositions and methods of using said CRISPR/Cas9-based system related compositions for altering gene expression and genome engineering. Also disclosed herein are compositions and methods of using said compositions for altering gene expression and genome engineering in muscle, such as skeletal muscle and cardiac muscle.
    Type: Grant
    Filed: May 29, 2018
    Date of Patent: August 18, 2020
    Assignee: Duke University
    Inventors: Charles A. Gersbach, Isaac B. Hilton, Pablo Perez-Pinera, Ami M. Kabadi, Pratiksha I. Thakore, David G. Ousterout, Joshua B. Black
  • Patent number: 10711285
    Abstract: The invention provides for delivery, 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. 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: Grant
    Filed: December 17, 2015
    Date of Patent: July 14, 2020
    Assignees: THE BROAD INSTITUTE, INC., MASSACHUSETTS INSTITUTE OF TECHNOLOGY, PRESIDENT AND FELLOWS OF HARVARD COLLEGE
    Inventors: Feng Zhang, Patrick Hsu, Chie-yu Lin, Fei Ran
  • Patent number: 10701911
    Abstract: The present invention relates generally to genetically modified non-human animals and immunodeficient non-human animals characterized by restored complement-dependent cytotoxicity, as well as methods and compositions for assessment of therapeutic antibodies in the genetically modified immunodeficient non-human animals. In specific aspects, the present invention relates to immunodeficient non-obese diabetic (NOD), A/J, A/He, AKR, DBA/2, NZB/BIN, B10.D2/oSn and other mouse strains genetically modified to restore complement-dependent cytotoxicity which is lacking in the unmodified immunodeficient mice. In further specific aspects, the present invention relates to NOD.Cg-Prkdcscid IL2rgtm1Wjl/SzJ (NSG), NOD.Cg-Rag1tm1Mom IL2rgtm1Wjl/SzJ (NRG) and NOD.Cg-Prkdcscid IL2rgtm1Sug/JicTAc (NOG) mice genetically modified to restore complement-dependent cytotoxicity which is lacking in unmodified NSG, NRG and NOG mice.
    Type: Grant
    Filed: June 16, 2016
    Date of Patent: July 7, 2020
    Assignees: The Jackson Laboratory, University of Massachusetts
    Inventors: Leonard D. Shultz, Mohit Kumar Verma, Dale L. Greiner, Michael A. Brehm
  • Patent number: 10550372
    Abstract: The invention provides for systems, methods, and compositions for altering expression of target gene sequences and related gene products. Provided are structural information on the Cas protein of the CRISPR-Cas system, use of this information in generating modified components of the CRISPR complex, vectors and vector systems which encode one or more components or modified components of a CRISPR complex, as well as methods for the design and use of such vectors and components. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for utilizing the CRISPR-Cas system. In particular the present invention comprehends optimized functional CRISPR-Cas enzyme systems.
    Type: Grant
    Filed: June 10, 2016
    Date of Patent: February 4, 2020
    Assignees: The Broad Institute, Inc., Massachusetts Institute of Technology, University of Tokyo, President and Fellows of Harvard College
    Inventors: Silvana Konermann, Alexandro Trevino, Mark Brigham, Fei Ran, Patrick Hsu, Chie-yu Lin, Osamu Nureki, Hiroshi Nishimasu, Ryuichiro Ishitani, Feng Zhang
  • Patent number: 10531648
    Abstract: A genetically modified mouse is provided that comprises a conditional Acvr1 allele that comprises a mutated exon that, upon induction, converts to a mutant exon phenotype, wherein the mutant exon phenotype includes ectopic bone formation. Mice comprising a mutant Acvr1 exon 5 in antisense orientation, flanked by site-specific recombinase recognition sites, are provided, wherein the mice further comprise a site-specific recombinase that recognizes the site-specific recombinase recognitions sites, wherein the recombinase is induced upon exposure of the mouse to tamoxifen. Upon exposure to tamoxifen, the recombinase is expressed and acts on the RRS-flanked mutant exon 5 and places the mutant exon 5 in sense orientation and deletes the wild-type exon.
    Type: Grant
    Filed: October 30, 2018
    Date of Patent: January 14, 2020
    Assignee: REGENERON PHARMACEUTICALS, INC.
    Inventors: Aris N. Economides, Sarah Jane Hatsell
  • Patent number: 10525082
    Abstract: Disclosed are methods of making a genetically modified immune cell for modifying a tumor microenvironment (TME) and methods of modifying a tumor microenvironment (TME). In some embodiments, the method can include delivering a first vector to an immune cell, wherein the first vector comprises a nucleic acid encoding a protein that induces T-cell proliferation, promotes persistence and activation of endogenous or adoptively transferred NK or T cells and/or induces production of an interleukin, an interferon, a PD-1 checkpoint binding protein, HMGB1, MyD88, a cytokine or a chemokine. Methods of modulating the suppression of the immune response in a tumor microenvironment, minimizing the proliferation of tumor and suppressive cells, and increasing the efficiency of an anti-cancer therapy, anti-infection therapy, antibacterial therapy, anti-viral therapy, or anti-tumoral therapy are also provided.
    Type: Grant
    Filed: September 7, 2016
    Date of Patent: January 7, 2020
    Assignee: Seattle Children's Hospital
    Inventors: Courtney Crane, Michael Jensen, Kara White Moyes, Nicole Lieberman
  • Patent number: 10519454
    Abstract: The disclosure provided herewith relates to a Campylobacter jejuni CRISPR/CAS system-derived RGEN and a use thereof.
    Type: Grant
    Filed: January 31, 2017
    Date of Patent: December 31, 2019
    Assignee: TOOLGEN INCORPORATED
    Inventors: Eun Ji Kim, Seok Joong Kim
  • Patent number: 10508152
    Abstract: The present invention relates generally to the field of RNA Control Devices and/or destabilizing elements (DE) combined with Chimeric Antigen Receptors (CARs) in eukaryotic cells. The present invention also relates to split CARs (Side-CARs) in eukaryotic cells. More specifically, the present invention relates to DEs, RNA Control Devices, and/or side-CARs combined with Chimeric Antigen Receptors to make small molecule actuatable CAR polypeptides. The present invention also relates to DE-CARs, Smart CARs (Smart=small molecule actuatable RNA trigger), Smart-DE-CARs, and/or Side-CARs for use in the treatment of disease.
    Type: Grant
    Filed: September 18, 2017
    Date of Patent: December 17, 2019
    Assignee: Chimera Bioengineering, Inc.
    Inventors: Benjamin Wang, Gusti Zeiner
  • Patent number: 10470444
    Abstract: A genetically modified mouse is provided that comprises a conditional Acvr1 allele that comprises a mutated exon that, upon induction, converts to a mutant exon phenotype, wherein the mutant exon phenotype includes ectopic bone formation. Mice comprising a mutant Acvr1 exon 5 in antisense orientation, flanked by site-specific recombinase recognition sites, are provided, wherein the mice further comprise a site-specific recombinase that recognizes the site-specific recombinase recognitions sites, wherein the recombinase is induced upon exposure of the mouse to tamoxifen. Upon exposure to tamoxifen, the recombinase is expressed and acts on the RRS-flanked mutant exon 5 and places the mutant exon 5 in sense orientation and deletes the wild-type exon.
    Type: Grant
    Filed: October 30, 2018
    Date of Patent: November 12, 2019
    Assignee: REGENERON PHARMACEUTICALS, INC.
    Inventors: Aris N. Economides, Sarah Jane Hatsell
  • Patent number: 10448621
    Abstract: A genetically modified mouse is provided that comprises a conditional Acvr1 allele that comprises a mutated exon that, upon induction, converts to a mutant exon phenotype, wherein the mutant exon phenotype includes ectopic bone formation. Mice comprising a mutant Acvr1 exon 5 in antisense orientation, flanked by site-specific recombinase recognition sites, are provided, wherein the mice further comprise a site-specific recombinase that recognizes the site-specific recombinase recognitions sites, wherein the recombinase is induced upon exposure of the mouse to tamoxifen. Upon exposure to tamoxifen, the recombinase is expressed and acts on the RRS-flanked mutant exon 5 and places the mutant exon 5 in sense orientation and deletes the wild-type exon.
    Type: Grant
    Filed: May 18, 2018
    Date of Patent: October 22, 2019
    Assignee: REGENERON PHARMACEUTICALS, INC.
    Inventors: Aris N. Economides, Sarah Jane Hatsell
  • Patent number: 10435699
    Abstract: Compositions and methods are provided for stacking multiple independent transgenic loci into the genome of a plant. Compositions include plants, seeds or plant cells comprising at least one transgenic target site and at least one genomic locus of interest integrated at different genomic sites within a genomic window. Plant breeding techniques can be employed such that the transgenic target site and the genomic locus of interest can be bred together. In this way, multiple independent transgene integrations can be generated within a genomic window to create a complex trait locus. The complex trait locus is designed such that the transgenic target sites and/or genomic loci of interest can segregate independently of each other, thus providing the benefit of altering a complex trait locus by breeding-in and breeding-away specific elements. Various methods can also be employed to modify the target sites such that they contain a variety of polynucleotides of interest.
    Type: Grant
    Filed: January 24, 2013
    Date of Patent: October 8, 2019
    Assignees: E I DU PONT DE NEMOURS AND COMPANY, PIONEER HI-BRED INTERNATIONAL, INC.
    Inventors: Saverio Carl Falco, Michael W Lassner, Zhongsen Li, Christopher Jay Scelonge
  • Patent number: 10392633
    Abstract: Nucleic acid constructs and methods for rendering modifications to a genome are provided, wherein the modifications comprise null alleles, conditional alleles and null alleles comprising COINs. Multifunctional alleles (MFA) are provided, as well as methods for making them, which afford the ability in a single targeting to introduce an allele that can be used to generate a null allele, a conditional allele, or an allele that is a null allele and that further includes a COIN. MFAs comprise pairs of cognate recombinase recognition sites, an actuating sequence and/or a drug selection cassette, and a nucleotide sequence of interest, and a COIN, wherein upon action of a recombinase a conditional allele with a COIN is formed. In a further embodiment, action of a second recombinase forms an allele that contains only a COIN in sense orientation. In a further embodiment, action by a third recombinase forms an allele that contains only the actuating sequence in sense orientation.
    Type: Grant
    Filed: July 12, 2013
    Date of Patent: August 27, 2019
    Assignee: REGENERON PHARMACEUTICALS, INC.
    Inventors: Aris N. Economides, Andrew J. Murphy, Peter Matthew Lengyel, Peter H. A. Yang
  • Patent number: 10323073
    Abstract: Methods and products (e.g., gRNAs, recombinant fusion proteins, frataxin targeting systems, compositions and kits) are described for increasing frataxin expression/levels in a cell, as well as uses of such methods and products, for example for the treatment of Friedreich ataxia in a subject suffering therefrom.
    Type: Grant
    Filed: March 20, 2015
    Date of Patent: June 18, 2019
    Assignee: UNIVERSITÉ LAVAL
    Inventors: Jacques P. Tremblay, Pierre Chapdelaine, Joël Rousseau
  • Patent number: 10081817
    Abstract: The present invention provides a method for obtaining site-specific recombination in a eukaryotic cell, the method comprising providing a eukaryotic cell that comprises a first recombination attachment site and a second recombination attachment site; contacting the first and second recombination attachment sites with a prokaryotic recombinase polypeptide, resulting in recombination between the recombination attachment sites, wherein the recombinase polypeptide can mediate recombination between the first and second recombination attachment sites, the first recombination attachment site is a phage genomic recombination attachment site (attP) or a bacterial genomic recombination attachment site (attB), the second recombination site is attB or attP, and the recombinase is selected from the group consisting of a Listeria monocytogenes phage recombinase, a Streptococcus pyogenes phage recombinase, a Bacillus subtilis phage recombinase, a Mycobacterium tuberculosis phage recombinase and a Mycobacterium smegmatis pha
    Type: Grant
    Filed: October 31, 2016
    Date of Patent: September 25, 2018
    Assignee: Intrexon Corporation
    Inventor: Malla Padidam
  • Patent number: 10030245
    Abstract: Methods for producing in a plant a complex transgenic trait locus comprising at least two altered target sequences in a genomic region of interest are disclosed. The methods involve the use of two or more double-strand-break-inducing agents, each of which can cause a double-strand break in a target sequence in the genomic region of interest which results in an alteration in the target sequence. Also disclosed are complex transgenic trait loci in plants. A complex transgenic trait locus comprises at least two altered target sequences that are genetically linked to a polynucleotide of interest. Plants, plant cells, plant parts, and seeds comprising one or more complex transgenic trait loci are also disclosed.
    Type: Grant
    Filed: March 22, 2012
    Date of Patent: July 24, 2018
    Assignees: E I DU PONT DE NEMOURS AND COMPANY, PIONEER HI-BRED INTERNATIONAL, INC.
    Inventors: Michael Lassner, Derek Jantz, James Jefferson Smith, Mark Cigan, Carl Falco, Huirong Gao, Zhongsen Li, Zhan-Bin Liu, Sergei Svitashev
  • Patent number: 9777064
    Abstract: The present invention relates generally to the field of RNA Control Devices and/or destabilizing elements (DE) combined with Chimeric Antigen Receptors (CARs) in eukaryotic cells. The present invention also relates to split CARs (Side-CARs) in eukaryotic cells. More specifically, the present invention relates to DEs, RNA Control Devices, and/or side-CARs combined with Chimeric Antigen Receptors to make small molecule actuatable CAR polypeptides. The present invention also relates to DE-CARs, Smart CARs (Smart=small molecule actuatable RNA trigger), Smart-DE-CARs, and/or Side-CARs for use in the treatment of disease.
    Type: Grant
    Filed: March 15, 2016
    Date of Patent: October 3, 2017
    Assignee: Chimera Bioengineering, Inc.
    Inventors: Benjamin Wang, Gusti Zeiner
  • Patent number: 9572837
    Abstract: The present disclosure relates to compositions and methods for reducing immune tolerance associated with CAR T cell therapy. Embodiments of the present disclosure include isolated nucleic acid sequence comprising a nucleic acid sequence that encodes modified programmed cell death protein 1 (PD-1) and a nucleic acid sequence that encodes chimeric antigen receptor (CAR).
    Type: Grant
    Filed: April 7, 2016
    Date of Patent: February 21, 2017
    Assignee: Innovative Cellular Therapeutics CO., LTD.
    Inventor: Zhao Wu
  • Patent number: 9510569
    Abstract: A genetically modified mouse is provided that comprises a conditional Acvr1 allele that comprises a mutated exon that, upon induction, converts to a mutant exon phenotype, wherein the mutant exon phenotype includes ectopic bone formation. Mice comprising a mutant Acvr1 exon 5 in antisense orientation, flanked by site-specific recombinase recognition sites, are provided, wherein the mice further comprise a site-specific recombinase that recognizes the site-specific recombinase recognitions sites, wherein the recombinase is induced upon exposure of the mouse to tamoxifen. Upon exposure to tamoxifen, the recombinase is expressed and acts on the RRS-flanked mutant exon 5 and places the mutant exon 5 in sense orientation and deletes the wild-type exon.
    Type: Grant
    Filed: March 12, 2014
    Date of Patent: December 6, 2016
    Assignee: Regeneron Pharmaceuticals, Inc.
    Inventors: Aris N. Economides, Sarah Jane Hatsell
  • Patent number: 9222105
    Abstract: Nucleases and methods of using these nucleases for modification of an HPRT locus and for increasing the frequency of gene modification at a targeted locus and clones and for generating animals.
    Type: Grant
    Filed: October 25, 2012
    Date of Patent: December 29, 2015
    Assignees: Sangamo BioSciences, Inc., The Regents of the University of California
    Inventors: Gregory J. Cost, Michael C. Holmes, Noriyuki Kasahara, Josee Laganiere, Jeffrey C. Miller, David Paschon, Edward J. Rebar, Fyodor Urnov, Lei Zhang
  • Patent number: 9163232
    Abstract: The invention relates to a method for manufacturing a recombinant polyclonal protein composition, in particular a recombinant polyclonal antibody composition. The method comprises obtaining a collection of cells transfected with a library of variant nucleic acid sequences, wherein each call in the collection is transfected with and capable of expressing one member of the library, which encodes a distinct member of a polyclonal protein that binds a particular antigen and which is located at the same single site in the genome of individual cells in said collection, wherein said nucleic acid sequence is not naturally associated with said cell in the collection. The cells are cultured under suitable conditions for expression of the polyclonal protein, which is obtained from the cells or culture supernatant. The present method is suitable for manufacturing recombinant polyclonal antibodies, thereby making available a superior replacement of plasma-derived therapeutic immunoglobulin products.
    Type: Grant
    Filed: December 19, 2008
    Date of Patent: October 20, 2015
    Assignee: Symphogen A/S
    Inventors: John S. Haurum, Finn C. Wiberg, Vincent W. Coljee, Jacqueline Sharon, Chiou-Ying Yang
  • Publication number: 20150140665
    Abstract: Methods for inserting a polynucleotide sequence into the genome of a human cell are provided. The present methods result in insertion of a polynucleotide sequence of interest into the H11 locus in the genome of a human cell. Also provided are nucleic acids that include sequences for integrating a polynucleotide sequence of interest into the H11 locus in the genome of a human cell. A transgenic human cell including site specific recombination sites at the H11 locus is also disclosed.
    Type: Application
    Filed: November 12, 2014
    Publication date: May 21, 2015
    Inventors: Michele Pamela Calos, Ruby Yanru Tsai, Fangfang Zhu, Matthew Gamboa, Alfonso P. Farruggio, Simon Hippenmeyer, Bosiljka Tasic, Birgitt Schüle
  • Publication number: 20150140664
    Abstract: Methods of simultaneously excising large nucleic acid sequences from a target nucleic acid and inserting large foreign nucleic sequences into the target nucleic acid sequence using DNA binding protein nucleases are described.
    Type: Application
    Filed: June 30, 2014
    Publication date: May 21, 2015
    Inventors: Susan M. BYRNE, George M. CHURCH
  • Patent number: 9034650
    Abstract: The present invention provides a method for obtaining site-specific recombination in a eukaryotic cell, the method comprising providing a eukaryotic cell that comprises a first recombination attachment site and a second recombination attachment site; contacting the first and second recombination attachment sites with a prokaryotic recombinase polypeptide, resulting in recombination between the recombination attachment sites, wherein the recombinase polypeptide can mediate recombination between the first and second recombination attachment sites, the first recombination attachment site is a phage genomic recombination attachment site (attP) or a bacterial genomic recombination attachment site (attB), the second recombination site is attB or attP, and the recombinase is selected from the group consisting of a Listeria monocytogenes phage recombinase, a Streptococcus pyogenes phage recombinase, a Bacillus subtilis phage recombinase, a Mycobacterium tuberculosis phage recombinase and a Mycobacterium smegmatis pha
    Type: Grant
    Filed: February 2, 2005
    Date of Patent: May 19, 2015
    Assignee: Intrexon Corporation
    Inventor: Malla Padidam
  • Publication number: 20150128300
    Abstract: The disclosure provides methods and compositions for generating conditional knock-out alleles using donor constructs together with sequence-specific nucleases to generate conditional knock-out alleles. Specifically, the donor construct comprises a 5? homology region, a 5? recombinase recognition site, a donor sequence, a 3? recombinase recognition site, and a 3? homology region. Further disclosed are the donor sequences each comprises a target sequence having at least one neutral mutation. Different sequence-specific nucleases can be used with the donor constructs are further disclosed.
    Type: Application
    Filed: June 12, 2013
    Publication date: May 7, 2015
    Inventors: Soren Warming, Keith R. Anderson
  • Publication number: 20150125958
    Abstract: Disclosed herein are methods for controlling stem cell differentiation through the introduction of transgenes having Xic, Tsix, or Xite sequences to block differentiation and the removal of the transgenes to allow differentiation. Also disclosed are small RNA molecules and methods for using the small RNA molecules to control stem cell differentiation. Also disclosed are stem cells genetically modified by the introduction of Xic, Tsix, or Xite sequences.
    Type: Application
    Filed: October 6, 2014
    Publication date: May 7, 2015
    Inventor: JEANNIE T. LEE
  • Publication number: 20150118755
    Abstract: The disclosure relates to a method of reprogramming one or more somatic cells, e.g., partially differentiated or fully/terminally differentiated somatic cells, to a less differentiated state, e.g., a pluripotent or multipotent state. In further embodiments the invention also relates to reprogrammed somatic cells produced by methods of the invention, to uses of said cells, and to methods for identifying agents useful for reprogramming somatic cells.
    Type: Application
    Filed: August 29, 2014
    Publication date: April 30, 2015
    Inventors: Rudolf Jaenisch, Yaqub Hanna, Marius Wernig, Christopher J. Lengner, Alexander Meissner, Oliver Tobias Brambrink, G. Grant Welstead, Ruth Foreman
  • Publication number: 20150110762
    Abstract: The present disclosure is in the field of genome engineering, particularly targeted modification of the genome of a cell.
    Type: Application
    Filed: October 16, 2014
    Publication date: April 23, 2015
    Inventors: Michael C. Holmes, Jianbin Wang
  • Publication number: 20150079681
    Abstract: 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: Application
    Filed: October 24, 2014
    Publication date: March 19, 2015
    Inventor: Feng Zhang
  • Publication number: 20150079064
    Abstract: Disclosed herein are transcription activator-like effector nuclease (TALEN)-related compositions and methods of using said TALENs for correcting mutant genes.
    Type: Application
    Filed: April 26, 2013
    Publication date: March 19, 2015
    Applicant: Duke University
    Inventors: Charles Gersbach, David Ousterout
  • Publication number: 20150079680
    Abstract: The invention relates to an approach for introducing one or more desired insertions and/or deletions of known sizes into one or more predefined locations in a nucleic acid (eg, in a cell or organism genome). They developed techniques to do this either in a sequential fashion or by inserting a discrete DNA fragment of defined size into the genome precisely in a predefined location or carrying out a discrete deletion of a defined size at a precise location. The technique is based on the observation that DNA single-stranded breaks are preferentially repaired through the HDR pathway, and this reduces the chances of indels (eg, produced by NHEJ) in the present invention and thus is more efficient than prior art techniques. The invention also provides sequential insertion and/or deletions using single- or double-stranded DNA cutting.
    Type: Application
    Filed: September 18, 2014
    Publication date: March 19, 2015
    Inventors: Allan Bradley, Hanif Ali, E-Chiang Lee
  • Publication number: 20150071899
    Abstract: Some aspects of this disclosure provide compositions, methods, and kits for improving the specificity of RNA-programmable endonucleases, such as Cas9. Also provided are variants of Cas9, e.g., Cas9 dimers and fusion proteins, engineered to have improved specificity for cleaving nucleic acid targets. Also provided are compositions, methods, and kits for site-specific nucleic acid modification using Cas9 fusion proteins (e.g., nuclease-inactivated Cas9 fused to a nuclease catalytic domain). Such Cas9 variants are useful in clinical and research settings involving site-specific modification of DNA, for example, genomic modifications.
    Type: Application
    Filed: June 30, 2014
    Publication date: March 12, 2015
    Applicant: President and Fellows of Harvard College
    Inventors: David R. Liu, John Paul Guilinger, David B. Thompson
  • Publication number: 20150072368
    Abstract: Novel transcription units that may be used in expression vectors. The transcription unit allow antibodies to be produced whose gain in productivity is not linked to a particular antigenic target antibody and therefore by extrapolation to a given recombinant protein, nor linked to the culture medium.
    Type: Application
    Filed: February 8, 2013
    Publication date: March 12, 2015
    Applicant: LABORATOIRE FRANCAIS DU FRACTIONNEMENT ET DES BIOTECHNOLOGIES
    Inventors: Alexandre Fontayne, François Coutard
  • Publication number: 20150071889
    Abstract: Disclosed herein are methods, compositions, and kits for high efficiency, site-specific genomic editing of cells.
    Type: Application
    Filed: September 12, 2014
    Publication date: March 12, 2015
    Inventors: Kiran Musunuru, Chad A. Cowan, Derrick J. Rossi
  • Publication number: 20150071898
    Abstract: Some aspects of this disclosure provide compositions, methods, and kits for improving the specificity of RNA-programmable endonucleases, such as Cas9. Also provided are variants of Cas9, e.g., Cas9 dimers and fusion proteins, engineered to have improved specificity for cleaving nucleic acid targets. Also provided are compositions, methods, and kits for site-specific recombination, using Cas9 fusion proteins (e.g., nuclease-inactivated Cas9 fused to a recombinase catalytic domain). Such Cas9 variants are useful in clinical and research settings involving site-specific modification of DNA, for example, genomic modifications.
    Type: Application
    Filed: June 30, 2014
    Publication date: March 12, 2015
    Applicant: President and Fellows of Harvard College
    Inventors: David R. Liu, John Paul Guilinger, David B. Thompson
  • Publication number: 20150071900
    Abstract: Some aspects of this disclosure provide compositions, methods, systems, and kits for controlling the activity and/or improving the specificity of RNA-programmable endonucleases, such as Cas9. For example, provided are guide RNAs (gRNAs) that are engineered to exist in an “on” or “off” state, which control the binding and hence cleavage activity of RNA-programmable endonucleases.
    Type: Application
    Filed: July 8, 2014
    Publication date: March 12, 2015
    Applicant: President and Fellows of Harvard College
    Inventors: David R. Liu, Johnny Hao Hu
  • Publication number: 20150067898
    Abstract: Methods, uses, and animals for introgression of alleles between animals, including SNPs. One embodiment involves introducing a targeted targeting endonuclease system and a HDR template into a cell with a mismatch in the binding of the targeting endonuclease and the targeted site.
    Type: Application
    Filed: April 28, 2014
    Publication date: March 5, 2015
    Inventors: Scott C. Fahrenkrug, Daniel F. Carlson
  • Publication number: 20150064789
    Abstract: 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: Application
    Filed: August 28, 2014
    Publication date: March 5, 2015
    Inventors: David Paschon, Lei Zhang
  • Publication number: 20150059010
    Abstract: 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: Application
    Filed: August 20, 2014
    Publication date: February 26, 2015
    Inventors: Andrew Mark Cigan, Phillip A. Patten, Joshua K. Young
  • Publication number: 20150056705
    Abstract: Methods and compositions for genetic alteration of cells are provided.
    Type: Application
    Filed: May 15, 2014
    Publication date: February 26, 2015
    Inventors: Anthony Conway, Gregory J. Cost, Russell Dekelver, Edward J. Rebar, Andreas Reik, Fyodor Urnov, Jianbin Wang, H. Steve Zhang
  • Publication number: 20150050739
    Abstract: The present invention relates to a method for carrying out recombination at a target locus.
    Type: Application
    Filed: March 12, 2013
    Publication date: February 19, 2015
    Inventors: Noel Nicolaas Maria Elisabeth Van Peij, Martina Beishuizen, Yvonne Johannes Odilia Arendsen
  • Publication number: 20150050699
    Abstract: 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: Application
    Filed: March 20, 2013
    Publication date: February 19, 2015
    Inventors: Virginijus Siksnys, Giedrius Gasiunas, Tautvydad Karvelis, Arvydas Lubys, Lolita Zaliauskiene, Monika Glemzaite, Anja Smith
  • Publication number: 20150044772
    Abstract: 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: Application
    Filed: August 8, 2014
    Publication date: February 12, 2015
    Inventor: Guojun Zhao
  • Publication number: 20150037892
    Abstract: The present invention relates to a method for carrying out recombination at a target locus.
    Type: Application
    Filed: March 12, 2013
    Publication date: February 5, 2015
    Inventors: Nathalie Wiessenhaan, Catharina Petronella Antonia Maria Kolen, Bernard Meijrink, Viktor Marius Boer, Johannes Andries Roubos, Yvonne Johannes Odilia Arendsen
  • Patent number: 8945884
    Abstract: 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: Grant
    Filed: May 13, 2013
    Date of Patent: February 3, 2015
    Assignee: Life Technologies Corporation
    Inventors: Jonathan Chesnut, John Carrino, Louis Leong, Knut Madden, Martin Gleeson, James Fan, Michael Brasch, David Cheo, James Hartley, Devon Byrd, Gary Temple
  • Publication number: 20150031133
    Abstract: Methods of genome engineering in cells using a TALEN lacking repeat sequences or Cas9 is provided.
    Type: Application
    Filed: June 30, 2014
    Publication date: January 29, 2015
    Inventors: George M. CHURCH, Luhan YANG, Marc Guell CARGOL, Joyce Lichi YANG