Abstract: The present invention discloses a method for modulating the production of a protein from a polynucleotide in a CHO cell by replacing at least one codon of the polynucleotide with a synonymous codon that has a higher or lower translation efficiency in the CHO cell than the codon it replaces, or by introducing into the CHO cell a polynucleotide that codes for an iso-tRNA which limits the rate of production of the polypeptide and which corresponds to a codon of the first polynucleotide. The present invention also discloses the use of a protein-encoding polynucleotide whose codon composition has been modified for enhanced production of the protein in CHO cells.

Abstract: An integration-defective retroviral vector transfer cassette lacking a functional polypurine tract (PPT) is provided. Also provided are isolated nucleic acids that include a heterologous nucleotide sequence, one or two retroviral long terminal repeats (LTRs), a packaging signal, a rev responsive element, and a eukaryotic promoter, wherein the nucleic acid lacks a functional PPT; vectors that include the disclosed isolated nucleic acids; recombinant retroviral particles and mRNAs thereof; retroviral vector kits; and methods for producing integration-defective vector particles, achieving gene expression of a nucleotide sequence of interest, and inserting a nucleotide sequence of interest into a host cell genome in a site-specific or non-specific manner.

Abstract: The present invention 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.

Abstract: Methods and vectors and kits are provided for producing chimeric nucleic acid constructs capable of producing dsRNA for silencing target nucleic acid sequences of interest using recombinational cloning.

Abstract: The present invention provides methods to site-specifically manipulate genomes by using hybrid recombinases. Hybrid recombinases comprise a modified catalytic domain from a unidirectional serine phage integrase, fused to a foreign DNA recognition domain.

Abstract: Provided are novel vectors and viral vectors capable of expressing exogenous gene or exogenous nucleic acid sequences in a target cell of interest, such as T cells, bone marrow cells, epithelial cells, liver cells and the like. The nucleic acid components of the vectors may include one or more native promoter/enhancer regions having modified sequence segments, one or more non-native promoter/enhancer or non-native promoter's gene or gene segment, and a native viral vector terminator or processing signal or segment thereof. The viral vectors comprise a virus or viral portion having on the surfaces or envelopes adsorption components, one for a packaging cell line and the other for delivery to a target cell. Other viral vectors provided by this invention have two components on their surfaces or envelopes, one of which is native to the virus and the other being non-native and capable of adsorbing to the target cell while being incapable of adsorbing to a native cell for the viral vector.

Abstract: A novel inbred maize line designated PHCK5 and seed, plants and plant parts thereof. Methods for producing a maize plant that comprise crossing inbred maize line PHCK5 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into PHCK5 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. Hybrid maize seed, plant or plant part produced by crossing the inbred line PHCK5 or a trait conversion of PHCK5 with another maize line. Inbred maize lines derived from inbred maize line PHCK5, methods for producing other inbred maize lines derived from inbred maize line PHCK5 and the inbred maize lines and their parts derived by the use of those methods.

Abstract: A method and oligonucleotides for targeted nucleotide exchange of a duplex DNA sequence, wherein the donor oligonucleotide contains at least one modified nucleotide which is a LNA having a higher binding affinity compared to naturally occurring A, C, T or G and/or binds stronger to a nucleotide in an opposite position in the first DNA sequence as compared to a naturally occurring nucleotide complementary to the nucleotide in the opposite position in the first DNA sequence.

Abstract: Yeast cells are mutagenized to obtain desirable mutants. Mutagenesis is mediated by a defective mismatch repair system which can be enhanced using conventional exogenously applied mutagens. Yeast cells with the defective mismatch repair system are hypermutable, but after selection of desired mutant yeast strains, they can be rendered genetically stable by restoring the mismatch repair system to proper functionality.

Abstract: The present invention generally relates to the fields of genetic engineering and antibody production. In particular, it relates to the generation of genetically modified vertebrate precursor lymphocytes that have the potential to differentiate into more mature lymphoid lineage cells, and to the use thereof for the production of any heterologous antibody or binding protein.

Abstract: A process of the production of a product of interest in an F1 seed obtained by a hybridization of a first and a second transgenic parental plant, said hybridization generating a genetic endowment in said F1 seed for said production by combining in said F1 seed first and second partial genetic endowments of said first and second transgenic parental plants, followed by isolating said product of interest from said F1 seed or a seedling thereof.

Abstract: Prokaryotic recombination systems have been adapted to function in eukaryotes in order to achieve one or more of the following: DNA site specific excision, translocation, integration and inversion. These recombination systems are identified as seven members of the small serine resolvase subfamily: CinH, ParA, Tn1721, Tn5053, Tn21, Tn402, and Tn501 and three members of the large serine resolvase subfamily: Bxb1, U153, and TP901-1. These recombination systems represent new tools for the genetic manipulation of eukaryotic genomes.

Abstract: A process of producing a transgenic multi-cellular plants or parts thereof expressing a trait of interest, said trait having a controlled distribution of said trait to progeny, wherein said process comprises (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 plant or cells thereof to generate progeny exhibiting said functional trait of interest due to binding between a protein or polypeptide encoded by said first heterologous nucleotide sequence and a protein or polypeptide encoded by said second heterologous nucleotide

Abstract: A method for conditionally knocking out and altering gene function and genetic sequences that can be used in such methods, for use in gene trapping and gene targeting. Specifically, the genetic sequence is a inducible gene silencer comprising: (a) a splice acceptor sequence; (b) an internal ribosomal entry site (IRES) sequence; (c) a nucleotide sequence coding for a reporter protein; (d) a polyadenylation sequence; and (e) a pair of oppositely oriented recombination site sequences, which cause single cycle inversions in the presence of a suitable recombinase enzyme, flanking elements (a) through (d).

Abstract: A method of creating a human pluripotent transgenic stem cell, wherein heterologous DNA is inserted into specific “hot-spots” in the genome where stable and high gene expression may occur, is disclosed. In one embodiment, the method comprises the steps of: (a) selecting a pluripotent stem cell line, and (b) inserting heterologous DNA at an insertion site selected from the group consisting of insertion site one and insertion site two to form a transgenic cell line. In another embodiment, the heterologous DNA is an exchange cassette and the transgenic cell line formed is a master cell line.

Abstract: A strategy for suppressing expression of one allele of an endogenous gene is provided comprising providing suppression effectors such as antisense nucleic acids able to bind to polymorphisms within or adjacent to a gene such that one allele of a gene is exclusively or preferentially suppressed and if required of a replacement gene can be introduced. The invention has the advantage that the same suppression strategy when directed to polymorphisms could be used to suppress, in principle, many mutations in a gene. This is particularly relevant when large numbers of mutations within a single gene cause disease pathology.

Abstract: The present invention provides a system for site-specific directed gene insertion of desired genes or foreign DNA into cellular genomes. The system includes novel vectors for integrating DNA into the genome of different hosts. Methods of using the vectors and transformed hosts are described.

Abstract: The invention provides an improved directed complementation method for generating a conditionally tumorigenic mouse cell. In a directed complementation method, the tumorigenicity of a conditionally tumorigenic mouse cell depends on either the expression of an inducible recombinant oncogene or the expression of a recombinant gene of interest that functionally complements an uninduced recombinant oncogene. The invention provides a method of producing a tumorigenic mouse cell containing an uninduced oncogene, a recombinant gene of interest that functionally complements the uninduced oncogene, and a Cre-ER system capable of excising the recombinant gene of interest. When the Cre-ER system is activated, the recombinant gene of interest is excised. From the effect on the mouse cell it is possible to determine whether the recombinant gene of interest is a tumor maintenance gene.

Abstract: A method for the production of fungus resistant transgenic plants, plant cells or plant tissue comprising the introduction of an Ab, rAb, rAb fragment or fusion or vector of the invention or the vectors of the composition of the invention into the genome of a plant, plant cell or plant cell tissue and a transgenic plant cell comprising stably integrated into the genome a polynucleotide or vector of the invention or the vectors of the composition of the invention.

Abstract: Artificial chromosomes, including ACes, that have been engineered to contain available sites for site-specific, recombination-directed integration of DNA of interest are provided. These artificial chromosomes permit tractable, efficient, rational engineering of the chromosome for a variety of applications.

Abstract: The invention provides cells and methods of circularizing linear DNA molecules. The cell is an isolated Escherichia coli cell which transiently expresses the Cre recombinase protein from an integrated Cre recombinase gene, and which is at least transiently repressed for RecBCD activity. The cells are used in a method of circularizing a linear DNA molecule comprising at least two loxP sites. The DNA molecule is introduced into the cells, and the linear DNA molecule is joined at said loxp sites.

Abstract: The present invention 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.

Abstract: The present invention provides methods of site-specifically integrating a polynucleotide sequence of interest in a genome of a eucaryotic cell, as well as, enzymes, polypeptides, and a variety of vector constructs useful therefore. In the method, a targeting construct comprises, for example, (i) a first recombination site and a polynucleotide sequence of interest, and (ii) a site-specific recombinase, which are introduced into the cell. The genome of the cell comprises a second recombination site. Recombination between the first and second recombination sites is facilitated by the site-specific recombinase. The invention describes compositions, vectors, and methods of use thereof, for the generation of transgenic cells, tissues, plants, and animals. The compositions, vectors, and methods of the present invention are also useful in gene therapy techniques.

Abstract: The present invention provides a method of achieving very high targeting efficiency by utilizing targeting vectors that utilize promoter-less selection cassettes and which are engineered to targeted into transcriptionally active loci. In particular, the invention provides a method for targeting promoter-less selection cassettes into transcriptionally active loci in stem cells or other eukaryotic cells with much greater efficiency than previously observed with other methods, thus reducing the number of drug-resistant clones to be screened or eliminating the need to screen for targeted cells altogether. The invention also encompasses the DNA targeting vectors, the targeted cells, as well as non-human organisms, especially mice, created from the targeted cells.

Abstract: The present invention provides for the nucleic acid sequences of plant centromeres. This will permit construction of stably inherited recombinant DNA constructs and minichromosomes which can serve as vectors for the construction of transgenic plant and animal cells.

Abstract: The disclosure relates generally to stem cell biology and more specifically to genetic manipulation of stem cells. Methods and compositions using recombinational cloning techniques are disclosed which allow the construction and insertion of complex genetic constructs into embryonic and adult stem cells and progenitor cells. The methods disclosed will allow the harvesting of adult stem cells pre-engineered with integration sites to facilitate early passage genetic modification.

Abstract: Systems, methods, compositions and apparatus relating to genome selection are disclosed.

Abstract: The present invention describes methods of identifying altered recombinases and compositions thereof, wherein at least one amino acid is different from a parent, wild-type recombinase and the altered recombinase has improved recombination efficiency towards wild-type and/or pseudo att site sequences relative to the parent, wild-type recombinase. The present invention also includes methods of modifying the genomes of cells using the altered recombinases, including methods of site-specifically integrating a polynucleotide sequence of interest in a genome of a eucaryotic cell.

Abstract: Serotonin neurons modulate most homeostatic Central Nervous System (CNS) functions while influencing the expression of behavioral traits such as mood, aggression and anxiety. Serotonin neuron dysfunction has been implicated in depression, addiction, autism and sudden infant death syndrome. This disclosure describes a straightforward, highly reproducible method for genetically accessing serotonin neurons and a sub-population of intestinal enterocytes, in vivo, using BAC-based transgenes that can be constructed using simple subcloning procedures. Compositions described herein include these transgenes and methods for making and using them to create transgenic mouse strains and identify serotonin and intestinal enterocytes without immunostaining.

Abstract: A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent ?-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebrate development.

Abstract: The present invention provides methods of site-specifically integrating a polynucleotide sequence of interest in a genome of a eucaryotic cell, as well as, enzymes, polypeptides, and a variety of vector constructs useful therefore. In the method, a targeting construct comprises, for example, (i) a first recombination site and a polynucleotide sequence of interest, and (ii) a site-specific recombinase, which are introduced into the cell. The genome of the cell comprises a second recombination site. Recombination between the first and second recombination sites is facilitated by the site-specific recombinase. The invention describes compositions, vectors, and methods of use thereof, for the generation of transgenic cells, tissues, plants, and animals. The compositions, vectors, and methods of the present invention are also useful in gene therapy techniques.

Abstract: The present disclosure provides methods for obtaining the targeted integration of a DNA molecule into the genome of a host cell using a recombinase. The methods disclosed herein can be used with a variety of host cells, including, for example, dicotyledonous and monocotyledonous plant cells. The present disclosure provides a method for effecting site-specific recombination of DNA within a plant cell, comprising: introducing into the plant cell a target nucleotide sequence comprising a first Int recognition site; introducing into the plant cell a donor nucleotide sequence comprising a second Int recognition site; and introducing into the plant cell an integrase or integrase complex.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: A novel inbred maize line designated PHENE and seed, plants and plant parts thereof. Methods for producing a maize plant that comprise crossing inbred maize line PHENE with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into PHENE through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. Hybrid maize seed, plant or plant part produced by crossing the inbred line PHENE or a trait conversion of PHENE with another maize line. Inbred maize lines derived from inbred maize line PHENE, methods for producing other inbred maize lines derived from inbred maize line PHENE and the inbred maize lines and their parts derived by the use of those methods.

Abstract: A novel inbred maize line designated PHAPT and seed, plants and plant parts thereof. Methods for producing a maize plant that comprise crossing inbred maize line PHAPT with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into PHAPT through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. Hybrid maize seed, plant or plant part produced by crossing the inbred line PHAPT or a trait conversion of PHAPT with another maize line. Inbred maize lines derived from inbred maize line PHAPT, methods for producing other inbred maize lines derived from inbred maize line PHAPT and the inbred maize lines and their parts derived by the use of those methods.

Abstract: The present invention provides methods for inducing insulin gene expression in cultured pancreas cells, the method comprising contacting a culture of endocrine pancreas cells expressing a PDX-1 gene with a GLP-1 receptor agonist, wherein the cells have been cultured under conditions such that the cells are in contact with other cells in the culture, thereby inducing insulin gene expression in the ceils. The invention also provides methods of treating a diabetic human subject using the methods of the invention.

Abstract: The present invention relates to a recombinant vector comprising MJ1 gene coding an integrase derived from Enterococcus temperate bacteriophage ? FC1 and an integration method using the vector. More particularly, the present invention relates to a recombinant vector comprising MJ1 gene which can make a site-specific integration in the human cell independently without other factors but not cause an excision and an integration method using the vector. Therefore, the present invention can be very useful in gene therapy in mammalian animal.

Abstract: A novel inbred maize line designated PHEHG and seed, plants and plant parts thereof. Methods for producing a maize plant that comprise crossing inbred maize line PHEHG with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into PHEHG through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. Hybrid maize seed, plant or plant part produced by crossing the inbred line PHEHG or a trait conversion of PHEHG with another maize line. Inbred maize lines derived from inbred maize line PHEHG, methods for producing other inbred maize lines derived from inbred maize line PHEHG and the inbred maize lines and their parts derived by the use of those methods.

Abstract: A novel inbred maize line designated PHAR1 and seed, plants and plant parts thereof. Methods for producing a maize plant that comprise crossing inbred maize line PHAR1 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into PHAR1 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. Hybrid maize seed, plant or plant part produced by crossing the inbred line PHAR1 or a trait conversion of PHAR1 with another maize line. Inbred maize lines derived from inbred maize line PHAR1, methods for producing other inbred maize lines derived from inbred maize line PHAR1 and the inbred maize lines and their parts derived by the use of those methods.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: A novel hybrid maize variety designated 33Y74 and seed, plants and plant parts thereof, produced by crossing Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 33Y74 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 33Y74 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 33Y74, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 33Y74. This invention further relates to methods for producing maize lines derived from hybrid maize variety 33Y74 and to the maize lines derived by the use of those methods.

Abstract: A method of assembling large DNA fragments in a chromosome using site specific recombinases and alternating excisionases. The method may be performed in vitro or in vivo, but larger assemblies are possible when the assembly is performed in vivo. For an in vivo assembly, the cell must be engineered to contain the desired recombinases, each in an inducible construct so that the desired recombinase can be expressed at the correct time with the correct choice of inducing agent.

Abstract: Disclosed is a method of making transgenic plants. Heterologous DNA is first introduced into a donor plant, plant cell or protoplast, and then moved from the donor to a recipient plant, plant cell or protoplast unaccompanied by any native genomic DNA of the donor. The donor and recipient are chosen that produce unstable progeny or demonstrate preferential segregation or sorting out. The DNA may be inserted randomly or at specific locations in the genome of the recipient plant. Also disclosed are transgenic plants produced by the methods, and plant progeny, plant parts and seeds and seed parts from the plants.

Abstract: The present invention provides methods for the conditional or regulated expression of a site-specific recombinase using split intein-mediated protein splicing. This enhances the temporal and tissue-specificity of trait gene expression and allows for fine tuning of expression specificity.

Abstract: The present invention provides for the nucleic acid sequences of plant centromeres. This will permit construction of stably inherited recombinant DNA constructs and minichromosomes which can serve as vectors for the construction of transgenic plant and animal cells.

Abstract: Compositions and methods for introducing a DNA of interest into a genomic target site are provided. In particular, the methods and compositions involve the use of a combination of target sites for two site specific recombinases and expression of a chimeric recombinase with dual target site specificity. Thus, the compositions comprise novel site-specific recombinases with specificities to multiple target sites, and nucleotide sequences and expression cassettes encoding these recombinases or target sites. The methods involve transforming a eukaryotic cell having target sites for the novel recombinase with a DNA of interest that is flanked by corresponding target sites. Expression of the recombinase results in integration of the DNA of interest into the genome of the cell. The compositions and methods of the invention have use in the construction of stably transformed eukaryotic cells, and in particular, plant cells.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: A novel hybrid maize variety designated 33G58 and seed, plants and plant parts thereof, produced by crossing Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 33G58 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 33G58 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 33G58, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 33G58. This invention further relates to methods for producing maize lines derived from hybrid maize variety 33G58 and to the maize lines derived by the use of those methods.

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: A novel hybrid maize variety designated 32B81 and seed, plants and plant parts thereof, produced by crossing Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 32B81 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 32B81 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 32B81, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 32B81. This invention further relates to methods for producing maize lines derived from hybrid maize variety 32B81 and to the maize lines derived by the use of those methods.