598 results about "Gene targets" patented technology

Gene targeting is a technique to introduce genetic change into one or more specific locations in the genome of a cell. For example, gene targeting can introduce genetic change by modifying, repairing, attenuating or inactivating a target gene or other chromosomal DNA. In one aspect, this disclosure relates to methods and compositions for gene targeting with high efficiency in a cell. This disclosure also relates to methods of treating or preventing a genetic disease in an individual in need thereof. Further disclosed are chimeric nucleases and vectors encoding chimeric nucleases.

The invention relates to a gene targeting system, which comprises two parts such as a site-specific cleavage nuclease expression vector and a targeting vector, wherein the targeting vector contains 2-10 donor DNA fragments, 5' ends and 3' ends of every donor DNA fragment are respectively inserted into recognition sequences of the site-specific cleavage nuclease, the donor DNA comprises an upstream homologous arm, a downstream homologous arm and an exogenous DNA sequence positioned between the upstream homologous arm and the downstream homologous arm, and the site-specific cleavage nuclease expression vector is any one selected from an expression vector carrying zinc finger nuclease, a transcription activator-like effector nuclease expression vector, and a RNA-mediated nuclease RNA:Cas9 expression vector.

A method for producing a transgenic plant includes providing a nucleic acid molecule comprising at least two regions of nucleic acid sequence that lack sequence homology with genomic DNA of the plant cell, and at least two zinc finger nuclease recognition sites, wherein the at least two regions of nucleic acid sequence that lack sequence homology with genomic DNA of the plant cell flank the at least two zinc finger nuclease recognition sites. A plant cell or tissue having the nucleic acid molecule stably integrated into the genome of the plant cell is transformed. A plant is regenerated from the plant cell. Transgenic plants are produced by the method. Seeds are produced by the transgenic plants.

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 invention discloses directional knockout on sheep MSTN gene by applying the CRISPR/Cas9 technology, and a method for verifying the impact effect of directional knockout on skeletal muscle satellite cell differentiation. The concrete contents are as follows: firstly, target gene cloning; secondly, gRBA design and synthesis; thirdly, CRISPR/Cas9 gene knockout vector construction; fourthly, CRISPR/Cas9 gene knockout vector exogenous activity detection; fifthly, CRISPR/Cas9 gene knockout vector endogenous activity detection; sixthly, CRISPR/Cas9 gene knockout vector knockout effect detection. The method has the following advantages: firstly, the experimental period is shorter; secondly, the method is simple and practicable, has high repeatability, and can be performed in common laboratories; thirdly, active vector constructed through the method is small in toxic and side effects, and can be applied to the preparation and production of transgenic animals; fourthly, the method is small in non-specific shear, and obviously improves the gene targeting knockout efficiency; fifthly, the method is high in applicability.

The invention belongs to the field of gene engineering and discloses a target sequence recognized by a CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 (CRISPR-associated protein 9) system stemming from streptococcus thermophilus. The target sequence is shown as the n-20th position of any one of SEQ ID NO: 1-88, wherein n ranges from 1 to 5. The invention further discloses sgRNA (single guide ribonucleic acid) with a sequence being 5'-recognition sequence-recruitment Cas9 sequence-3' and a coding DNA (deoxyribonucleic acid) molecule thereof, and a DNA sequence corresponding to the recognition sequence is as same as the target sequence. The invention further discloses the CRISPR-Cas9 system comprising Cas9 and the sgRNA and/or Cas9-carried coding sequence and an sgRNA coding sequence vector. The invention further discloses application of the CRISPR-Cas9 system to CCR5 gene editing and HIV (human immunodeficiency virus) infection drug preparation. CCR5 gene editing can be achieved to protect cells from HIV infection.

The invention provides a method for precisely editing a genome DNA sequence. A TALEN, CRISPR/Cas9 and other gene targeting technologies and homologous recombination technologies are utilized in a combined mode for precisely editing the genome DNA sequence. According to the method, the technical problem that in the prior art, a specific sequence cannot be deleted is solved, the specific base sequence can be seamlessly inserted or deleted in a fixed point mode, and seamless free modification of the genome DNA sequence of eukaryocyte is achieved for the first time.

The invention belongs to the field of gene engineering and discloses a target sequence identified by a CRISPR-Cas9 system from neisseria meningitidis. The target sequence is as shown in the nth-24th sites of optional one of SEQ ID No. 1-20, and n=1-9. The invention further discloses sgRNA and coding DNA molecules thereof, the sequence of the sgRNA is 5'-identificiation sequence-sequence recruiting Cas9 protein-3', and the DNA sequence corresponding to the identification sequence is identical with the target sequence. The invention also discloses the CRISPR-Cas9 system which comprises the Cas9 protein, the sgRNA and/or a coding sequence carrying the Cas9 protein and a vector of the coding sequence of the sgRNA. The invention also discloses application of the CRISPR-Cas9 system to editing of CCR5 genes and preparation of medicine for HIV infection. By the CRISPR-Cas9 system, the CCR5 genes can be edited, and cells cannot be infected by HIV.

The invention discloses a site-specific insertional inactivation method mediated by agrobacterium tumefaciens and CRISPR/Cas9. Endogenous snRNA promoter of Ustilago scitaminea (u6 promoter of Ustilago scitaminea) is used to drive sgRNA expression cassette. CRISPR/Cas9 system integrates with Ti plasmid of agrobacterium tumefaciens to construct the Ustilago scitaminea site-specific insertional inactivation system mediated by agrobacterium tumefaciens with hygromycin as a selection marker. Specific sequence of the objective gene is cloned into sgRNA expression cassette for transformation of Ustilago scitaminea basidiospores, thus the mobile DNA fragment is exactly inserted into the objective gene sequence of Ustilago scitaminea, achieving the goal of damage to gene function. The invention provides an important tool for study of Ustilago scitaminea functional genomics. The system has the advantages of high efficiency and accuracy, and is convenient to use to conduct gene function researches in Ustilago scitaminea.

The invention belongs to the field of gene engineering and discloses a target sequence identified by a CRISPR-Cas9 system from neisseria meningitidis. The target sequence is as shown in the nth-24th sites of optional one of SEQ ID No. 1-15, and n=1-9. The invention further discloses sgRNA and coding DNA molecules thereof, the sequence of the sgRNA is 5'-identificiation sequence-sequence recruiting Cas9 protein-3', and the DNA sequence corresponding to the identification sequence is identical with the target sequence. The invention also discloses the CRISPR-Cas9 system which comprises the Cas9 protein, the sgRNA and/or a coding sequence carrying the Cas9 protein and a vector of the coding sequence of the sgRNA. The invention also discloses application of the CRISPR-Cas9 system to editing of CXCR4 genes and preparation of medicine for HIV infection. By the CRISPR-Cas9 system, the CXCR4 genes can be edited, and cells cannot be infected by HIV.

The invention provides sgRNA and a gene vector for inhibiting bladder cancer by targeting human lncRNA-UCA1 and an application of the sgRNA; specifically, sgRNA sequences of an lncRNA-UCA1 gene and a PD-1 gene suitable for CRISPR-Cas9 targeted splicing are designed; by transforming plasmids of specific splicing lncRNA-UCA1 gene and CRISPR-Cas9 nuclease gene into human bladder cancer cells, the expression of the lncRNA-UCA1 gene drops so as to inhibit the growth of tumor cells; and the plasmid, together with a human PD-1 gene targeted knockout vector, is transformed into a humanized mouse model of bladder cancer transplanted tumor, so as to obviously inhibit the growth of tumors. According to the invention, the vector is simple in preparing steps, the sgRNA is good in targeting property and the CRISPR-Cas9 is high in knockout efficiency.

The invention provides a CRISPR-Cas9 system and an application thereof in treating breast cancer diseases, wherein by providing a plurality of sgRNAs (single guide RNAs), the efficient knockout of an RecQL4 gene can be achieved. In comparison with ZFNs and TALENs complex expression structures, siRNA is low in using complexity and efficiency; a Cas9 expression structure in the CRISPR-Cas9 system is fixed; for different genes, system building can be completed by interpolating recognition sequences into the sgRNA expression structure, so that operations are simplified and cost is reduced; and the system is applicable to mammal gene targeting in a large scale.

The invention discloses an artificial fixed-point rice dense and erect panicle (DEP1) gene mutant body and application thereof. The rice DEP1 gene mutant body as well as a corresponding allele is obtained by modifying a 5th exon of a DEP1 gene by adopting a CRISPR/Cas9 gene targeting modification technology, so that a base of the 5th exon is replaced, lost and inserted. The artificial fixed-point rice DEP1 gene mutant body and the application thereof disclosed by the invention have the benefits that the artificial mutation is performed on the 5th exon of the rice DEP1 gene by adopting the CRISPR/Cas9, and the mutant body capable of obviously improving the rice yield and the allele are screened; the DEP1 gene mutant body disclosed by the invention as well as the corresponding allele can improve the yield of rice plants by 13-51 percent and is even superior to a natural mutation type.

The invention discloses a preparation method of a liver cancer detection panel. The preparation method comprises the following steps: screening liver cancer information and related genes thereof, thusobtaining a gene set; selecting a gene target zone; designing a panel probe; synthesizing an RNA (Ribonucleic Acid) single-stranded probe, thus obtaining the gene detection panel. The gene detectionpanel designed by the invention can be used for diagnosis, typing and prognosis of liver cancer, has wide gene coverage degree and strong transferability and is suitable for newly-developed detectionplatforms.

The invention discloses a CYP3A4 gene SNP detection specific primer, a liquid-phase chip and a method; and the liquid-phase chip comprises wild-type and mutant ASPE primers which are respectively designed for each type of mutation points, microspheres which are respectively coated with specific anti-tag sequences, and the primer which is used for amplifying a CYP3A4 gene target sequence with CYP3A4*1B, CYP3A4*2, CYP3A4*3, CYP3A4*4, CYP3A4*5, CYP3A4*6, CYP3A4*15, CYP3A4*17, CYP3A4*18 and/or CYP3A4*19SNP sites. The CYP3A4 gene SNP detection liquid-phase chip has very good signal-noise ratio, and a designed probe and the anti-tag sequences essentially have no cross reaction. The designed ASPE primers have very good specificity and can accurately distinguish all types of mutation points. The detection method has simple steps and ten types of SNP sites can be detected at one step, so that the operation is convenient, thereby preventing a plurality of uncertain factors in a plurality of operation processes, and greatly improving the detection accuracy rate.

RTP801 represents a unique gene target for hypoxia-inducible factor-1 (HIF-1). Down-regulation of the mTOR pathway activity by hypoxia requires de novo mRNA synthesis and correlates with increased expression of RTP801. The present invention relates to screening systems utilizing RTP801 and/or RTP801 interactors and/or RTP801 biological activity, to drug candidates identified by such screening systems, and to the use of such drug candidates in the treatment of various disorders.

The invention discloses primers and probes for detecting human EGFR gene mutations as well as a use method, relating to the detection of gene mutation. The method of the invention comprises the following steps: (1) providing the primers and the probes; (2) processing the sample to be detected and extracting a template; (3) preparing a reaction system of fluorescent PCR amplification mutation gene sequence; (4) amplifying the mutable gene target sequence by the primers and the probes in the step (1); (5) detecting the fluorescence intensity of FAM and HEX or ROX of the reaction system as the judgment standard of results by adopting hybridization of a dual ring probes and an amplification product. The primers, the probes and the method of the invention can simultaneously detect 29 deletion mutations of EGFR gene, and features high sensitivity, strong specificity, strong selective capacity and fast detection speed demonstrated by completing the whole detection process in only 90 minutes.

The invention discloses an effective Cas13a-based anti-dengue virus nucleic acid target and application thereof. The invention provides a CRISPR-Cas13a system for inhibiting dengue virus, wherein theCRISPR-Cas13a system comprises a Cas13a protein and a crRNA corresponding to an NS3 gene target of the dengue virus, or a complex formed by the Cas13a protein and crRNA; and NS3 gene targets of the dengue virus are 4657th-4685th nucleotide sequences of the NS3 gene. In the invention, a novel anti-dengue virus method is found, is different from a traditional anti-virus method, directly targets a viral nucleic acid, specifically degrades a target gene, and makes the virus lose the replication ability; therefore, the effective Cas13a-based anti-dengue virus nucleic acid target has characteristicsof high efficiency, high specificity and programmability, is not easy to produce drug resistance generated by traditional antiviral drugs, and may become a novel antiviral drug in the future.