498 results about "Vector system" patented technology

Provided are retroviral vector genomes and vector systems comprising the genomes. In particular, a retroviral vector genome comprising two or more NOIs, operably linked by one or more Internal Ribosome Entry Site(s); a lentiviral vector genome comprising two or more NOIs suitable for treating a neurodegenerative disorder; and a lentiviral vector genome which encodes tyrosine hydroxylase, GTP-cyclohydrolase I and, optionally, Aromatic Amino Acid Dopa Decarboxylase are provided.

The present invention relates to retroviral vector genomes and to vector systems comprising such genomes. In particular the present invention relates to a retroviral vector genome comprising two or more NOIs operably linked by one or more Internal Ribosome Entry Site(s); a lentiviral vector genome comprising two or more NOIs suitable for treating a neurodegenerative disorder; and a lentiviral vector genome which encodes tyrosine hydroxylase, GTP-cyclohydrolase I and optionally Aromatic Amino Acid Dopa Decarboxylase.

The present invention relates to a retroviral vector system comprising a packaging cell line that synthesize the core and enzymatic proteins of MLV virus from one or more gag and pol containing constructs and the envelope of MMTV virus from the same or an independent env containing construct, and to pseudotyped retroviral particles produced by culturing said retroviral vector system transfected with a MLV based retroviral vector, and isolation of said pseudotyped retroviral particles.

The present invention provides a dual expression vector, and methods for its use, for the expression and secretion of a full-length polypeptide of interest in eukaryotic cells, and a soluble domain or fragment of the polypeptide in bacteria. When expressed in bacteria, transcription from a bacterial promoter within a first intron and termination at the stop codon in a second intron results in expression of a fragment of the polypeptide, e.g., a Fab fragment, whereas in mammalian cells, splicing removes the bacterial regulatory sequences located in the two introns and generates the mammalian signal sequence, allowing expression of the full-length polypeptide, e.g., IgG heavy or light chain polypeptide. The dual expression vector system of the invention can be used to select and screen for new monoclonal antibodies, as well as to optimize monoclonal antibodies for binding to antigenic molecules of interest.

The present invention provides multiply deficient adenoviral vectors and complementing cell lines. Also provided are recombinants of the multiply deficient adenoviral vectors and a therapeutic method, particularly relating to gene therapy, vaccination, and the like, involving the use of such recombinants.

A translation enhancer-driven positive feedback vector system is disclosed which is designed to facilitate identification of a Translational Enhancer Element (TEE) and to provide a means for overexpression of gene products. The system exploits both transcriptional and translational approaches to control the expression levels of genes and / or gene products. Methods are also disclosed for screening libraries of random nucleotide sequences to identify translational elements and for overproduction of proteins, which have uses in both research and industrial environments.

The invention relates to a clustered regularly interspaced short palindromic repeats (CRISPR) / Cas guide RNA (gRNA) comprising a targeting domain that is complementary to human genomic Epidermal Growth Factor Receptor (EGFR) DNA, and a vector system including one or more packaged vector(s) including: (a) a first regulatory element operably linked to a gRNA, and (b) a second regulatory element operably linked to a nucleic acid encoding a Cas protein. Also disclosed are methods of altering a nucleic acid sequence encoding EGFR in a cell including contacting the cell with a vector system, methods of treating lung cancer, and methods of selectively inducing apoptosis in a cell including administering a gRNA to the cell.

A translation enhancer-driven positive feedback vector system is disclosed which is designed to facilitate identification of a Translational Enhancer Element (TEE) and to provide a means for overexpression of gene products. The system exploits both transcriptional and translational approaches to control the expression levels of genes and / or gene products. Methods are also disclosed for screening libraries of random nucleotide sequences to identify translational elements and for overproduction of proteins, which have uses in both research and industrial environments.

This invention provides a method for treating a subject having a tumor and a method for inhibiting angiogenesis in a subject, both comprising administering to the subject an effective amount of a composition of matter comprising the extracellular domain of a Notch receptor protein operably affixed to a half-life-increasing moiety. This invention also provides a composition of matter comprising the extracellular domain of Notch4 receptor protein operably affixed to a half-life-increasing moiety. This invention further provides an article of manufacture. Finally, this invention provides a replicable vector which encodes a polypeptide comprising the extracellular domain of a Notch receptor protein operably affixed to a half-life-increasing moiety, a host vector system which comprises such replicable vector and a method of producing such polypeptide.

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.

The present invention generally relates to libraries, kits, methods, applications and screens used in functional genomics that focus on gene function in a cell and that may use vector systems and other aspects related to Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas systems and components thereof. The present invention also relates to rules for making potent single guide RNAs (sgRNAs) for use in CRISPR-Cas systems. Provided are genomic libraries and genome wide libraries, kits, methods of knocking out in parallel every gene in the genome, methods of selecting individual cell knock outs that survive under a selective pressure, methods of identifying the genetic basis of one or more medical symptoms exhibited by a patient, and methods for designing a genome-scale sgRNA library.

The present invention provides bacterial vectors and fusion proteins containing a TTSS polypeptide, compositions of such fusion proteins including polynucleotides, and methods of delivering one or more genes into a target cell that involve contacting the cell with a composition that includes such a fusion protein. Compositions and methods of gene delivery that involve a bacterium and a TAT, Antp, or HSV VP22 polypeptide are also disclosed. The invention also concerns methods of delivering one or more genes into a target cell utilizing a bacterium capable of becoming internalized within the cell, wherein the bacterium includes one or more genes targeted for delivery to the cell, a gene encoding an RNA polymerase, and a gene that causes lysis of the bacterium.

The present invention provides a dual expression vector, and methods for its use, for the expression and secretion of a full-length polypeptide of interest in eukaryotic cells, and a soluble domain or fragment of the polypeptide in bacteria. When expressed in bacteria, transcription from a bacterial promoter within a first intron and termination at the stop codon in a second intron results in expression of a fragment of the polypeptide, e.g., a Fab fragment, whereas in mammalian cells, splicing removes the bacterial regulatory sequences located in the two introns and generates the mammalian signal sequence, allowing expression of the full-length polypeptide, e.g., IgG heavy or light chain polypeptide. The dual expression vector system of the invention can be used to select and screen for new monoclonal antibodies, as well as to optimize monoclonal antibodies for binding to antigenic molecules of interest.

The present invention provides an improved method for achieving efficient transcription and translation of modified transgene constructs in vector systems. The vector may be a lentiviral vector. Such a method facilitates the production of viral vector genomes with intact functional transgene sequences allowing stable integration of a transgene-containing viral vector genome into the germline of an animal such as a transgenic avian. The subsequent expression of the transgene results in a recombinant protein product being produced, which, in the case of a transgenic avian can result in the targeted production of the protein into the egg of the transgenic bird.

The invention belongs to the field of gene engineering, and particularly relates to a method for specifically knocking out CCR5 genes of human bodies through CRISPR / SaCas9 and sgRNA used for specifically targeting CCR5 genes. The invention provides the method for specifically knocking out the CCR5 genes of the human bodies through CRISPR / SaCas9 and sgRNA used for specifically targeting the CCR5 genes and relates to a CRISPR / SaCas9 recombinant lentivirus and adeno-associated virus vector system and application. CRISPR / SaCas9 recombinant lentivirus and adeno-associated virus vectors can express SaCas9-RNA aiming at the CCR5 target spot of one same gene of the human bodies and rhesus monkeys, therefore, the application range is wider, and the efficiency is higher. The preparation method is easy to construct, the targeting efficiency is high, and a novel technology is provided for the gene therapy of HIV.

The invention relates to a CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9-system-based Saccharomyces cerevisiae genome concurrent multiplex edition vector and application thereof. Cas9 protein expression cassettes and a plurality of sgRNA (small guide ribonucleic acid) scaffold expression cassettes are integrated into the vector to obtain a vector-Cas9-sgRNA scaffold, and a plurality of sgRNA segments designed and synthesized by using ADE1 as a target are integrated into the vector-Cas9-sgRNA scaffold, thereby obtaining the Saccharomyces cerevisiae genome concurrent multiplex edition vector. By adopting a simple substance granule single-copy vector system, one plasmid is utilized to simultaneously express the Cas9 protein and three sgRNA scaffolds, so that three target genes can be edited simultaneously; and thus, the multigene edition operation is very simple. Besides, the target gene edition system only needs one-step component and conversion, and thus, is simple to operate.

The invention relates to a method and apparatus that uses an R-tree's vector data as a mechanism for query and retrieval of raster data in conjunction with or instead of the vector data. The invention relates to a method of storing, indexing and retrieving raster data comprising: storing raster data, storing vector data, and indexing said raster data and said vector data with same index information.

The invention provides new strains of the Modified Vaccinia Virus Ankara (MVA) that have a strongly reduced virulence for most mammals, especially humans, but nevertheless grows in cells of a continuous cell line approved for the production of a therapeutic agent such as a vaccine. The invention also provides a method for producing said adapted MVA strains. The adapted MVA can be used e.g. for parenteral immunization, as a vector system, or in the active or inactivated from as an adjuvant or as a regulator of the unspecific components of the immune system.

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 deliver systems and tissues or organ which are targeted as sites for delivery. Also 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 ceils to ensure enhanced specificity for target recognition and avoidance of toxicity and to edit or modify a target site in a genomic locus of interest to alter or improve the status of a disease or a condition.

Two mechanisms are provided for improving the expression of Factor IX in gene therapy vectors. The first is the use of a specific Factor IX polynucleotide coding sequence designed for optimal expression. The second is the use of transcriptional regulatory regions minimized in size so that they can be used to express Factor IX, as well as any other gene of interest, in a size-constrained environment such as in a self complementary gene therapy vector system.

Methods are provided for generating immune responses utilizing alphavirus-based vector systems.

The invention provides a combined navigation and positioning method of a small underwater robot, which comprises the following steps: fusing measurement data of various navigation facilities by means of a strong tracking unscented Kalman filter; using position vectors and heading angles under a navigation system as well as speed vectors and acceleration vectors under a vector system as state vectors of the filter; using the strong tracking unscented Kalman filter to implement autonomous navigation and data filtering of the underwater robot; based on horizontal position information output by a GPS (global positioning system) receiver, using the strong tracking unscented Kalman filter to implement autonomous correction and data filtering of the underwater robot; and implementing the switching between two different measurement equations (i.e. overwater and underwater measurement equations) based on the significance bit of the output signal of the GPS receiver. The invention can fuse the position, depth and attitude information of the small underwater robot measured by various navigation facilities, and can implement the autonomous navigation and autonomous correction of the small underwater robot under the interference of sea current or sea wave.

A host-vector system that uses the RNA-based copy number control mechanism of ColE1-type plasmids for regulating the expression of a marker gene allows for antibiotic-free selection of plasmids and is useful for production of plasmid DNA and recombinant proteins.

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.

The invention discloses CRISPR-Cas9 targeting knockout of human breast cancer cell RASSF2 gene and specific sgRNA of RASSF2 gene. According to the method, the sgRNA of specific targeting RASSF2 gene is obtained, wherein the base sequence of the sgRNA is represented by SEQ ID NO.1; construction of the sgRNA of RASSF2 gene into a lentiviral vector system is carried out, wherein the lentiviral vectorsystem contains Cas9 protein; and at last, human breast cancer cell MDA-MB-231 is infected with the CRISPR-Cas9 lentivirus containing the sgRNA so as to obtain a cell strain with obviously reduced RASSF2 protein expression level. The operation and the steps are simple; the sgRNA targeting performance is excellent; cutting efficiency on RASSF2 gene is high; the constructed CRISPR-Cas9 lentiviral vector system is high in knockout efficiency, and is capable of realizing specific knockout of RASSF2 gene to obtain human breast cancer cells without RASSF2 gene, so that powerful instrument is provided for study on the action mechanisms of RASSF2 in breast cancer celles.

The present invention provides an improved helper-dependent vector system for production of high capacity adenoviral cloning vectors. The invention makes use of the DNA size packaging constraints imposed on a pIX-defective Ad virion that prevent such virions from packaging DNA larger than approximately 35 kb. This constraint can be used to develop helper viruses that do not package their DNA. In one embodiment, the invention combines this methodology with the Cre-loxP helper-dependent system to decrease the quantity of contaminating helper virus in vector preparations. In another embodiment the invention is used for vector growth.

The invention discloses a CRISPR-Cas9 targeted and knockout human breast cancer cell SLC30A1 gene and the specific sgRNA of the gene. Firstly, sgRNA for specific targeting of the SLC30A1 gene is obtained, and a base sequence of the sgRNA is shown in SEQ ID NO.1; secondly, the sgRNA of the SLC30A1 gene is constructed to a lentivirus vector system containing Cas9 proteins; finally, CRISPR / Cas9 lentivirus containing the sgRNA is infected with a human breast cancer cell MDA-MB-231 to obtain a cell strain of which the SLC30A1 protein expression level is obviously reduced. The CRISPR-Cas9 targeted and knockout human breast cancer cell SLC30A1 gene and the specific sgRNA of the gene disclosed by the invention have the advantages of simple operation steps, good sgRNA targetability and high cuttingefficiency on the SLC30A1 gene; in addition, the constructed CRISPR / Cas9 lentivirus system has the advantage of high knockout efficiency and can specifically knock out the SLC30A1 gene so as to obtain the human breast cancer cell having the SLC30A1 gene knocked out; therefore, a powerful tool is provided for further researching an action mechanism of the SLC30A1 in breast cancer cells.

The invention discloses a CRISPR / Cas9 targeted knockout human colorectal carcinoma cell DEAF1 gene and specific sgRNA thereof. The CRISPR / Cas9 targeted knockout human colorectal carcinoma cell DEAF1 gene and the specific sgRNA thereof are characterized in that: firstly, sgRNA of a second exon of the specific targeted DEAF1 gene is obtained and the base sequence of the sgRNA is shown as SEQ ID NO.1; secondly, the sgRNA of the DEAF1 gene is constructed into a lentiviral vector system, which contains Cas9 protein; finally, the CRISPR / Cas9 lentivirus containing the sgRNA is infected with human colorectal carcinoma cell HT-29 cell, so that a cell strain of which DEAF1 protein expression level is obviously reduced is obtained. The CRISPR / Cas9 targeted knockout human colorectal carcinoma cell DEAF1 gene disclosed by the invention has the advantages of simple operation steps, good sgRNA target ability and high cutting efficiency for the DEAF1 gene; in addition, the constructed CRISPR / Cas9 lentiviral vector system has the advantage of high knockout efficiency and can specifically knock out the DEAF1 gene to obtain the human colorectal carcinoma cells knocking out the DEAF1 gene, and therebya powerful tool is provided for further studying an action mechanism of DEAF1 in the colorectal carcinoma cells.

The invention provides new strains of the Modified Vaccinia Virus Ankara (MVA) that have a strongly reduced virulence for most mammals, especially humans, but nevertheless grows in cells of a continuous cell line approved for the production of a therapeutic agent such as a vaccine. The invention also provides a method for producing said adapted MVA strains. The adapted MVA can be used e.g. for parenteral immunization, as a vector system, or in the active or inactivated from as an adjuvant or as a regulator of the unspecific components of the immune system.

The present invention discloses CRISPR / Cas9 targeted knockout human intestinal cancer cell RITA gene and specific sgRNA thereof. First, sgRNA specifically targeted to a second exon of the RITA gene isobtained, and the base sequence of the sgRNA is as shown in SEQ ID NO. 1; secondly, a sgRNA lentiviral vector system of the RITA gene is constructed, and the sgRNA lentiviral vector system contains Cas9 protein; and finally human intestinal cancer HT-29 cells are infected with CRISPR / Cas9 lentivirus containing the sgRNA to obtain a cell line which is significantly reduced in RITA protein expression level. The invention has the advantages of simple operation steps, good sgRNA targeting property, and high RITA gene cutting efficiency; furthermore, the constructed CRISPR / Cas9 lentivirus system has the advantage of high knock-out efficiency and can specifically knock out the RITA gene to obtain RITA gene-knockout human intestinal cancer cells, and a powerful tool for the further study of theaction mechanism of RITA in the intestinal cancer cells is provided.