Abstract: Methods of creating mutations in genomic exons by inserting introns into the genomic exons via homologous recombination. Also, methods are provided for introducing modifications into genomic exons by inserting introns into the genomic exons via homologous recombination such that a mature mRNA transcript produced from a genomic region of the genome comprising the genomic exon does not contain the modification are provided. The methods provide for a rapid method for introducing mutations and/or modifications of any type into a mammalian cell genome.

Abstract: Cells and methods of using these cells for expressing a transgene expressing a protein that is aberrantly expressed in a metabolic disorders from a safe harbor locus.

Abstract: Methods for the targeted integration of nucleotide sequences into a plant are provided. Transfer cassettes comprising nucleotide sequences of interest flanked by non-identical recombination sites are used to transform a plant comprising a target site. The target site contains at least a set of non-identical recombination sites corresponding to those on the transfer cassette. Exchange of the nucleotide sequences flanked by the recombination sites is effected by a recombinase.

Abstract: Disclosed herein are polypeptides, polynucleotides encoding, cells and organisms comprising novel DNA-binding domains, including TALE DNA-binding domains. Also disclosed are methods of using these novel DNA-binding domains for modulation of gene expression and/or genomic editing of endogenous cellular sequences.

Abstract: Disclosed herein are polypeptides, polynucleotides encoding, cells and organisms comprising novel DNA-binding domains, including TALE DNA-binding domains. Also disclosed are methods of using these novel DNA-binding domains for modulation of gene expression and/or genomic editing of endogenous cellular sequences.

Abstract: A transgenic cat with a phenotype characterized by the substantial absence of the major cat allergen, Fel d I. The phenotype is conferred in the transgenic cat by disrupting the coding sequence of the target gene with a specialized construct. The phenotype of the transgenic cat is transmissible to its offspring.

Abstract: ABSTRACT Methods, compositions, constructs, vectors, cell lines, and kits, for generating induced pluripotent stem cells by site-specific integration of pluripotency coding sequences with endonucleases for use in gene therapy, regenerative medicine, cell therapy or drug screening.

Abstract: Regulatory elements, specifically replicators and transgene constructs containing replicator nucleic acid sequences, are disclosed herein. Methods of using replicators and transgene constructs including replicators to inhibit, delay, or prevent gene silencing are also disclosed herein.

Abstract: Methods and compositions for modifying T-cells in which PD1 and/or CTLA-4 is repressed and/or inactivated using fusion proteins such as artificial transcription factors and nucleases.

Abstract: Disclosed are methods and cellular systems for the generation of predetermined or random alterations at specific genomic location. The invention provides a transient expression and reverse transcription system to generate single-stranded DNA sequences homologous to a target genomic sequence, which can be transported to the nucleus to alter the genetic information of the target genomic sequence. Also provided are cellular and molecular components that can be used to increase the efficiency of the targeted genomic modification process.

Abstract: Compositions and methods related to stromal antigen 2 (STAG2) and its role in diverse human cancers, including nucleic acids, polypeptides, vectors, cells and cell lines.

Abstract: Disclosed herein are methods and compositions for targeted cleavage of a genomic sequence, targeted alteration of a genomic sequence, and targeted recombination between a genomic region and an exogenous polynucleotide homologous to the genomic region. The compositions include fusion proteins comprising a cleavage domain (or cleavage half-domain) and an engineered zinc finger domain and polynucleotides encoding same. Methods for targeted cleavage include introduction of such fusion proteins, or polynucleotides encoding same, into a cell. Methods for targeted recombination additionally include introduction of an exogenous polynucleotide homologous to a genomic region into cells comprising the disclosed fusion proteins.

Abstract: Methods and compositions for a genetic disease are provided.

Abstract: The present disclosure provides a DNA-targeting RNA that comprises a targeting sequence and, together with a modifying polypeptide, provides for site-specific modification of a target DNA and/or a polypeptide associated with the target DNA. The present disclosure further provides site-specific modifying polypeptides. The present disclosure further provides methods of site-specific modification of a target DNA and/or a polypeptide associated with the target DNA The present disclosure provides methods of modulating transcription of a target nucleic acid in a target cell, generally involving contacting the target nucleic acid with an enzymatically inactive Cas9 polypeptide and a DNA-targeting RNA. Kits and compositions for carrying out the methods are also provided. The present disclosure provides genetically modified cells that produce Cas9; and Cas9 transgenic non-human multicellular organisms.

Abstract: In some embodiments the present disclosure provides a composition for targeted alteration of a DNA sequence and methods of altering the targeted DNA sequence using the composition. In some embodiments such a composition comprises a MiniVector comprising a nucleic acid sequence template for homology-directed repair, alteration, or replacement of the targeted DNA sequence within a cell in vivo or in vitro, where the MiniVector lacks both a bacterial origin of replication and an antibiotic selection gene, and wherein the MiniVector has a size up to about 2,500 base pairs.

Abstract: An object is to provide a Tol1 element transposase and a use thereof. Provided is a Tol1 element transposase containing (a) a protein having the amino acid sequence of SEQ ID No: 1 or (b) a protein having an amino acid sequence homologous to the amino acid sequence of SEQ ID NO: 1 and having an enzymatic activity for transferring Tol1 element. Further, provided is a polynucleotide encoding the transposase and an expression construct containing the polynucleotide therein. The present invention also provides a DNA introduction system including (a) a donor factor having such a structure that a desired DNA is inserted in a transposase gene-defected Tol1 element and (b) a helper factor containing the transposase or the polynucleotide.

Abstract: The invention relates to a set of genetic constructs which allow the efficient and reproducible introduction of a specific nucleotide sequence at a fixed position in the genome by generating a double strand break at a specific position in the genome using a meganuclease and so stimulating a homologous recombination event at this locus between the genomic site and a transfected donor sequence. The present invention also relates to methods using these constructs and to these materials in the form of a kit.

Abstract: The invention concerns the use of a mutagenic agent blocking DNA replication in the cell for inserting in vitro a nucleic acid of interest inside a predetermined nucleotide sequence present in a chromosome contained in a prokaryotic or eukaryotic cell, said nucleic acid of interest being, prior to its insertion, included in a DNA vector which replicates in said prokaryotic or eukaryotic host cell.

Abstract: A process of producing transgenic multi-cellular plants or parts thereof expressing a trait of interest that has a controlled distribution of said trait to progeny, comprising (i) producing a first plant or a cell thereof having in a first locus of a nuclear chromosome a first heterologous nucleotide sequence comprising a first fragment of a nucleotide sequence encoding said trait of interest, (ii) producing a second plant or a cell thereof having in a second locus of a nuclear chromosome homologous to said nuclear chromosome of step (i), a second heterologous nucleotide sequence comprising a second fragment of the nucleotide sequence encoding said trait of interest, and (iii) hybridising said first and said second plants or cells thereof to generate progeny exhibiting said functional trait of interest. Also disclosed is a process of producing hybrid seeds for agriculture.

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.

Abstract: Compositions and methods are provided for integrating one or more genes of interest into cellular DNA without substantially disrupting the expression of the gene at the locus of integration, i.e., the target locus. These compositions and methods are useful in any in vitro or in vivo application in which it is desirable to express a gene of interest in the same spatially and temporally restricted pattern as that of a gene at a target locus while maintaining the expression of the gene at the target locus, for example, to treat disease, in the production of genetically modified organisms in agriculture, in the large scale production of proteins by cells for therapeutic, diagnostic, or research purposes, in the induction of iPS cells for therapeutic, diagnostic, or research purposes, in biological research, etc. Reagents, devices and kits thereof that find use in practicing the subject methods are also provided.

Abstract: Disclosed are methods for suppressing or altering gene expression by locked nucleic acids that have high binding affinity for target sequences in genomic DNA. The present methods include a step of recombination to insert a foreign DNA fragment into a specific target site in a genome with greater efficiency than with current techniques. Using a protein to anchor a locked nucleic acid targeting construct at a specific site, DNA recombination between that site and foreign DNA is induced by employing the cell's innate repair machinery. As a result of this recombination, gene transcription can be effectively suppressed. The present methods work in all mammalian cells, and provide a rapid methodology to examine gene function. The foreign DNA can carry markers for ease of screening. The technique is applied in the production of stable cell lines with inserted or deleted genes.

Abstract: This invention is directed to a general method for the chronic treatment, potential cure, or prevention of various metabolic and related diseases in people, including diabetes, by modulating IRS2 activity in cells and tissues in the body. IRS1 and IRS2 are part of the insulin or insulin-like growth factor signaling pathway. By upregulating the levels or functional activity of IRS2, insulin is used more efficiently by the body to control nutrient levels. By upregulating IRS2 levels or functional activity in pancreatic ?-cells, glucose sensing and insulin secretion are enhanced.

Abstract: The invention discloses methods for the generation of chimaeric human-non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanised antibodies; compositions comprising said antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in said methods.

Abstract: Disclosed herein are methods and compositions for targeted integration of a exogenous sequence into a predetermined target site in a genome for use, for example, in protein expression and gene inactivation.

Abstract: Provided is a method for preparing B cells which produce a human-type antibody, comprising substituting an antibody gene of B cells with a human antibody gene, wherein the B cells are non-human vertebrate B cells capable of inducing or halting AID (activation induced cytidine deaminase) expression with the induction of the expression of an exogenous Cre recombinase gene through extracellular stimulation followed by the inversion of the direction of the exogenous AID gene by expressed Cre recombinase.

Abstract: The invention provides molecular switches which couple external signals to functionality and to methods of making and using the same. The switches according to the invention can be used, for example, to regulate gene transcription, target drug delivery to specific cells, transport drugs intracellularly, control drug release, provide conditionally active proteins, perform metabolic engineering, and modulate cell signaling pathways. Libraries comprising the switches and expression vectors and host cells for expressing the switches are also provided.

Abstract: Rationally-designed LAGLIDADG meganucleases and methods of making such meganucleases are provided. In addition, methods are provided for using the meganucleases to generate recombinant cells and organisms having a desired DNA sequence inserted into a limited number of loci within the genome, as well as methods of gene therapy, for treatment of pathogenic infections, and for in vitro applications in diagnostics and research.

Abstract: Rationally-designed LAGLIDADG meganucleases and methods of making such meganucleases are provided. In addition, methods are provided for using the meganucleases to generate recombinant cells and organisms having a desired DNA sequence inserted into a limited number of loci within the genome, as well as methods of gene therapy, for treatment of pathogenic infections, and for in vitro applications in diagnostics and research.

Abstract: Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3?-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.

Abstract: The present invention relates to a fusion protein having a TAL (transcription activator-like) effector (TALE) domain and a nucleotide cleavage domain, and more particularly, to the TAL effector nuclease comprising a TAL (transcription activator-like) effector (TALE) domain and a nucleotide cleavage domain, wherein the TALE domain includes one or more TALE-repeat modules, each of the TALE-repeat modules recognizing a single specific nucleic acid, and a use thereof.

Abstract: The present disclosure includes methods for manipulation of the genome, to products obtained or obtainable from such methods, and uses of these products.

Abstract: Disclosed herein are enhanced polypeptides, polynucleotides encoding these polypeptides, cells and organisms comprising novel DNA-binding domains, including TALE DNA-binding domains. Also disclosed are methods of using these novel DNA-binding domains for modulation of gene expression and/or genomic editing of endogenous cellular sequences.

Abstract: The present invention relates to methods of producing a reversibly immortalized cell, cells obtainable by the above method, eukaryotic cells comprising one or more nucleic acid(s) coding for one or more immortalizing protein(s) and methods of re-differentiating these cells.

Abstract: Methods and compositions for regulating HIV infection and/or replication in which an anti-HIV transgene is integrated into the genome of a cell using a nuclease.

Abstract: The invention relates to a method of generating junctional variability in the nucleotide sequence of a polynucleotide of interest present in an intrachromosomal substrate/context in a eukaryotic cell which is competent for canonical Non Homologous End Joining pathway (NHEJ) repair, involving the generation of double-strand break (DSB) in the DNA sequence of said polynucleotide, and involving the use of polymerase Terminal Deoxynucleotidyl Transferase (TdT) in conditions enabling said TdT to add Non-templated nucleotides (N nucleotides) before ligation through the canonical Non Homologous End Joining pathway (NHEJ) thereby allowing a mutagenic repair to take place at the DSB site.

Abstract: The present invention provides PiggyBac transposase proteins, nucleic acids encoding the same, compositions comprising the same, kits comprising the same, non-human transgenic animals comprising the same, and methods of using the same.

Abstract: The present invention relates to 1-Cre1 variants which can in particular recognise and cleave DNA targets which do not comprise the same nucleotides at positions ±6 and ±7 which are present in the wild type 1-Cre1 target. The present invention also relates to 1-Cre1 variants which can recognise and cleave targets which do not comprise the wild type nucleotides at positions ±4, ±5, ±6, ±7 and to 1-Cre1 variants with new specificity which can recognise and cleave targets which do not comprise the wild type nucleotides at positions ±4, ±5, ±6, ±7, ±8, ±9 and ±10.

Abstract: The present invention provides an isolated RNA molecule comprising: a) an alphavirus 5? replication recognition sequence, wherein at least one initiation codon has been removed from the 5? replication recognition sequence; b) a nucleotide sequence encoding an alphavirus structural protein; and c) an alphavirus 3? replication recognition sequence, with the proviso that the RNA molecule does not contain a promoter that directs transcription of the nucleotide sequence of (b), and wherein the alphavirus 5? and 3? replication recognition sequences of (a) and (c) direct replication of the RNA molecule in the presence of alphavirus non-structural proteins.

Abstract: Nucleases and methods of using these nucleases for alteration of a CFTR gene and generation of cells and animal models.

Abstract: The present invention provides methods and kits for editing specific chromosomal sequences in cells. In particular, targeting endonucleases and single-stranded nucleic acids are used to edit the chromosomal sequence.

Abstract: The present invention relates to a method of sequence specific recombination of DNA in eukaryotic cells utilizing att sequences from the bacteriophage lambda. A particular embodiment of the invention relates to a method further comprising performing the sequence specific recombination of DNA with an Int and a Xis factor. The present invention further relates to vectors containing each of these sequences and their use as medicaments.

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.

Abstract: The invention provides methods for reprogramming somatic cells to generate multipotent or pluripotent cells. Such methods are useful for a variety of purposes, including treating or preventing a medical condition in an individual. The invention further provides methods for identifying an agent that reprograms somatic cells to a less differentiated state.

Abstract: The present invention refers to methods for selectively recognizing a base pair in a DNA sequence by a polypeptide, to modified polypeptides which specifically recognize one or more base pairs in a DNA sequence and, to DNA which is modified so that it can be specifically recognized by a polypeptide and to uses of the polypeptide and DNA in specific DNA targeting as well as to methods of modulating expression of target genes in a cell.

Abstract: The present invention relates to nucleotide sequences, which are variants of att recombination sequences, involved in sequence-specific recombination of DNA in eukaryotic cells, whereby sequence specific recombination is performed by a bacteriophage lambda integrase Int. Such att recombination sequences being e.g. attP.b, attP.a, attL.a, attR.a and attR.b. The present invention further relates to a method of sequence-specific recombination of DNA in eukaryotic cells, comprising the introduction of a first DNA comprising a nucleotide sequence containing at least one recombination sequence into a cell, introducing a second DNA comprising a nucleotide sequence containing at least one further recombination sequence into a cell, and performing the sequence specific recombination by a bacteriophage lambda integrase Int, whereby at least one of said first or second DNAs is an att recombination sequence being e.g. attP.b, attP.a, attL.a, attR.a or attR.b.

Abstract: The present invention is directed to the use of the maize Ac/Ds transposable elements in vertebrates.

Abstract: The present invention provides methods for using endogenous transcriptional control systems to regulate the expression of heterologous protein(s). In particular, targeted genome editing is used to integrate a sequence encoding the heterologous protein(s) in-frame with an endogenous coding sequence such that the expression of the heterologous and endogenous sequences is regulated by the endogenous control system.

Abstract: A method for inducing a site-specific modification in the HPRT gene, for a non-therapeutic purpose, by contacting a DNA target sequence selected from the group consisting of the sequences SEQ ID NO: 1 to 14 thereby cleaving the DNA target with an I-CreI variant or single-chain derivative having at least one substitution in one of the two functional subdomains of the LAGLIDADG (SEQ ID NO: 153) core domain situated from positions 26 to 40 and 44 to 77 of I-CreI.

Abstract: An I-CreI variant which has at least two substitutions, one in each of the two functional subdomains of the LAGLIDADG (SEQ ID NO: 229) core domain situated from positions 26 to 40 and 44 to 77 of I-CreI, said variant being able to cleave a DNA target sequence from a xeroderma pigmentosum gene. Use of said variant and derived products for the prevention and the treatment of Xeroderma pigmentosum.