2182 results about "Gene mutation" patented technology

Disclosed herein are structure-based modelling methods for the preparation of acetohydroxy acid synthase (AHAS) variants, including those that exhibit selectively increased resistance to herbicides such as imidazoline herbicides and AHAS inhibiting herbicides. The invention encompasses isolated DNAs encoding such variants, vectors that include the DNAs, and methods for producing the variant polypeptides and herbicide resistant plants containing specific AHAS gene mutations. Methods for weed control in crops are also provided.

The invention discloses a wheat genome site-specific modification method which comprises the following steps: enabling wheat tissues to contain guide RNA (ribonucleic acid) and Cas9 nuclease; under the coaction of the guide RNA and Cas9 nuclease, cutting a double-chain target segment on the target gene; and by utilizing the DNA (deoxyribonucleic acid) restoration function of the wheat cells, finally implementing random insertion and / or random deletion on the target segment in the wheat target gene. The experiment proves that the method disclosed by the invention can successfully perform gene mutation on wheat.

Methods and vectors (both DNA and retroviral) are provided for the construction of a Library of mutated cells. The Library will preferably contain mutations in essentially all genes present in the genome of the cells. The nature of the Library and the vectors allow for methods of screening for mutations in specific genes, and for gathering nucleotide sequence data from each mutated gene to provide a database of tagged gene sequences. Such a database provides a means to access the individual mutant cell clones contained in the Library. The invention includes the described Library, methods of making the same, and vectors used to construct the Library. Methods are also provided for accessing individual parts of the Library either by sequence or by pooling and screening. The invention also provides for the generation of non-human transgenic animals which are mutant for specific genes as isolated and generated from the cells of the Library.

Efficient methods are disclosed for the high throughput identification of mutations in genes in members of mutagenized populations. The methods comprise DNA isolation, pooling, amplification, creation of libraries, high throughput sequencing of libraries, preferably by sequencing-by-synthesis technologies, identification of mutations and identification of the member of the population carrying the mutation and identification of the mutation.

The subject invention discloses a method for determining the prognosis and probable clinical course of a subject diagnosed with B-CLL. Specifically, the invention involves comparing CD38 expression in a biological sample from the subject containing B-CLL cells to a baseline level of CD38 expression, wherein an elevated level of CD38 expression in relation to the baseline level of CD38 expression may indicate poor prognosis or aggressive course of disease in the subject. Also disclosed is a method for determining whether the Ig V genes of the B-CLL cells of a B-CLL patient are mutated, comprising comparing CD38 expression in a biological sample from the subject containing B-CLL cells to a baseline level of CD38 expression, wherein a lower level of CD38 expression in relation to the baseline level indicates IG V gene mutation.

The present invention provides methods for enhancing discriminatory RNA silencing by RNA silencing agents. In particular, the invention provides methods for generating RNA silencing agents which can discriminate between target and non-target mRNAs that differ in sequence by only one nucleotide. Also provided are improved RNA silencing agents with enhanced discriminatory RNA silencing, e.g., single nucleotide discriminatory RNA silencing. The compositions and methods of the invention are useful in therapeutic strategies for treating genetic disorders associated with dominant, gain-of-function gene mutations.

Novel vectors are described that incorporate, inter alia, a novel 3' gene trap cassette which can be used to efficiently trap and identify previously unknown cellular genes. Vectors incorporating the described 3' gene trap cassette find particular application in gene discovery and in the production of mutated cells and animals.

Infectious cDNA clones of North American Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) are provided. Further provided are cDNA clones comprising genetic mutations. Also provided are vaccines comprising cDNAs, including genetically and immunologically marked vaccines for North American PRRSV.

The present invention provides methods for determining the level of protein activity in a cell by: (i) measuring abundances of cellular constituents in a cell in which the activity of a specific protein is to be determined so that a diagnostic profile is thus obtained; (ii) measuring abundances of cellular constituents that occur in a cell in response to perturbations in the activity of said protein to obtain response profiles and interpolating said response profiles to generate response curves; and (iii) determining a protein activity level at which the response profile extracted from the response curves best fits the measured diagnostic profile, according to some objective measure. In alternative embodiments, the present invention also provides methods for identifying individuals having genetic mutations or polymorphisms that disrupt protein activity, and methods for identifying drug activity in vivo by determining the activity levels of proteins which interact with said drugs.

The miR15 and miR16 micro RNA genes are located at 13q14 within a 30 kb region of loss characteristic of cells from certain cancers, such as cells from chronic lymphocytic leukemia or prostate cancer. Chronic lymphocytic leukemia or prostate cancer can be diagnosed by detecting a reduction in miR15 or miR16 gene copy number, by determining miR15 or miR16 gene mutational status, or by detecting a reduction in the RNA transcribed from these genes. The miR15 or miR16 gene products can inhibit the neoplastic or tumorigenic growth of cancers such as chronic lymphocytic leukemia or prostate cancer cells when administered to subjects suffering from these diseases.

The invention provides methods and compositions for detecting a mutation in a target gene in a sample of blood or a fraction thereof, including in certain examples, a fraction that includes circulating tumor DNA. The methods can include a tiling PCR reaction, for example a one-sided multiplex tiling reaction. Virtually any type of mutation can be detected with the methods and compositions. In certain embodiments, gene fusions are detected. Improved PCR methods, especially for performing nested multiplex PCR reactions are provided.

This invention relates generally to the field of cancer diagnostics. The invention further relates to the use of mutational load distribution analysis (MLDA) to examine changes in the distribution of genetic mutations incident to cancer.

Methods and vectors (both DNA and retroviral) are provided for the construction of a Library of mutated cells. The Library will preferably contain mutations in essentially all genes present in the genome of the cells. The nature of the Library and the vectors allow for methods of screening for mutations in specific genes, and for gathering nucleotide sequence data from each mutated gene to provide a database of tagged gene sequences. Such a database provides a means to access the individual mutant cell clones contained in the Library. The invention includes the described Library, methods of making the same, and vectors used to construct the Library. Methods are also provided for accessing individual parts of the Library either by sequence or by pooling and screening. The invention also provides for the generation of non-human transgenic animals which are mutant for specific genes as isolated and generated from the cells of the Library.

The invention discloses a method of detecting gene mutation based on Blocker primers and ARMS primers, and a kit. According to the invention, corresponding Blocker primers are designed according to an annealing temperature, the Blocker primers are combined with a wild-type template under the annealing temperature of the Blocker to block amplification of the wild type, and simultaneously an ARMS technology is used to reduce amplification of the wild-type background. The method integrates a Blocker enrichment technology and an ARMS fluorescent quantification PCR technology in one reaction system, and thus realizes enrichment and identification for the mutated genes for one step, reduces operation steps, and improves detection sensitivity and singularity.

Described is a method for treating an individual having a neurological disorder with an associated mutation or mutations in a gene encoding a lysosomal enzyme. Specifically, the individual is administered a specific pharmacological chaperone for the lysosomal enzyme which increases trafficking of the protein from the ER to the lysosome in neural cells, with or without concomitantly increasing enzyme activity in neural cells. Restoration of trafficking relieves cell stress and other toxicities associated with accumulation of mutant proteins. Restoration of enzyme activity relieves substrate accumulation and pathologies associated with lipid accumulation. In a specific embodiment, the neurological disorder is Parkinson's disease or parkinsonism which is associated with mutations in glucocerebrosidase.

The invention discloses a method for carrying out in-situ repairing on blood coagulation factor F8 / F9. The method comprises the following steps: in a target genome sequence, selecting the mutation sites of blood coagulation factor as the gene sites for in-situ repairing; designing the binding sites of nuclease of sgRNA sequence of a CRISPR / Cas system; designing a homologous recombinant repairing donor sequence for in-situ repairing; delivering nuclease protein and / or sgRNA and the nucleotide sequence of the homologous recombinant repairing donor to the gene sites of in-situ repairing by a delivering carrier; generating damages to the genome DNA by the nuclease on the gene in-situ repairing sites; and inserting the homologous recombinant repairing donor sequence into the gene in-situ repairing sites so as to repair the gene or supplement the expression of gene. The in-situ repairing of mutation sites of blood coagulation factor can be applied to the clinic, and the method has the advantages of precise induction, safer and controllable process, and definite target.

Gene targeting allows the deletion (knock out), the repair (rescuing) and the modification (gene mutation) of a selected gene and the functional analysis of any gene of interest. Targeting of nuclear genes has been a very inefficient process in most eukaryotes including plants and animals due to the dominance of illegitimate integration of the applied DNA into non-homologous regions of the genome. The present invention provides a method for increasing the ratio of homologous to non-homologous recombination of a polynucleotide into a host cell's DNA by suppressing non-homologous recombination. Surprisingly, the number of non-homologous recombination events can be reduced if the polynucleotide is applied as a purified single-stranded DNA, preferably coated with a single strand binding protein.

A method of genotyping women experiencing infertility for non-physical reasons in order to identify the presence of the rs4238001 and / or rs10846744 mutation of the SCARB1 gene and, upon identifying the presence of one or both genetic mutations, administering a tailored therapeutic regimen to restore fertility by either one or a combination of 1) mediating the flux of cholesterol resulting from the mutation by therapeutic use of the cholesterol medication probucol and / or other cholesterol altering medications, and / or 2) amplifying the presence of hormone progesterone by therapeutic use of progestational and progestin medications.

DNA amplification and hybridization are successively carried out in a reaction system containing primers for the DNA amplification and hybridization probes, followed by detecting the hybrid in the reaction solution by affinity chromatography, wherein at least one of the primers to be used in the DNA amplification is labeled with a first labeling agent so that the amplified DNA will be labeled with the first labeling agent, a hybridization probe is labeled with a second labeling agent and contained in a reaction solution for effecting the DNA amplification, the base sequence of the hybridization probe is designed not to inhibit the DNA amplification, and a hybrid is detected by affinity chromatography with the use of the first and second labeling agents.

Described herein are methods and compositions for treating an ER-related disease condition characterized by a mutation in the ESR1 gene by administering an estrogen receptor modulator. Also described herein are methods of treating hormone resistant-estrogen receptor (ER) positive breast cancers characterized by a mutation in the ESR1 gene by administering an estrogen receptor modulator.

The subject invention provides materials and methods wherein unique and advantageous combinations of gene mutations are used to direct carbon flow from sugars to a single product. The techniques of the subject invention can be used to obtain products from native pathways as well as from recombinant pathways. In preferred embodiments, the subject invention provides new materials and methods for the efficient production of acetate and pyruvic acid.

Plants are disclosed that contain a mutation in a delta-12 or delta-15 fatty acid desaturase gene. Preferred plants are rapeseed and sunflower plants. Plants carrying such mutant genes have altered fatty acid composition in seeds. In one embodiment, a plant contains a mutation in a region having the conserved motif His-Xaa-Xaa-His, found in delta-12 and delta-15 fatty acid desaturases. A preferred motif has the sequence His-Glu-Cys-Gly-His. A preferred mutation in this motif has the amino acid sequence His-Lys-Cys-Gly-His. Nucleic acid fragments are disclosed that comprise a sequence of at least 20 nucleotides from a mutant delta-12 or delta-15 fatty acid desaturase gene.

The invention provides an enrichment and analysis method for circulating tumor cells. The method mainly comprises the following steps: subjecting a biological fluid sample to red blood cell lysis treatment; adding immune beads specifically bonding with white cells, red cells and / or platelets for incubation; and filtering cell suspension by using a filter membrane and enriching circulating tumor cells. According to the invention, through integration and optimization of CTCs enrichment methods such as red blood cell lysis treatment, an immune bead method and membrane filtration, the enrichment effect of the circulating tumor cells is greatly improved; with the method, the removal rate of leukocytes is greater than 99.9%, and the enrichment rate of tumor cells is greater than 80%. Meanwhile, the invention also provides a kit using the enrichment method for the circulating tumor cells and discloses a device for enrichment of the circulating tumor cells. The invention also provides an analysis method for the enriched circulating tumor cells. With the analysis method, analysis of dynamic changes like the amount of tumor cells, gene mutation and gene expression profiles can be realized.

The invention discloses a recombinant vector and a non-GMO gene editing plant screening method. An initial carrier in the recombinant vector is a CRISPR / Cas9 carrier which contains sgRNA gene, Cas9 gene and screening label gene and is used for plant gene editing, the recombinant vector carries a BelRNAi expression element, and a hairpin RNA interference fragment interfering with bentazone resistance gene is generated through transcription of the BelRNAi expression element. By introducing the BelRNAi expression element to the initial carrier and conducting bentazone herbicide screening on future generation plants, offspring with target gene mutated are reserved, and it is also ensured that the mutated offspring do not contain transgenic fragments; screening method is cheaper, simpler and more effective.

The present invention provides methods of marker assisted selection for high oleic / low linolenic traits in canola and in other oil seed crop species, as well as isolated nucleic acids for use as molecular markers in such methods. In particular, molecular markers and Brassica nucleic acid corresponding to fad2 and fad3 gene mutations are disclosed. The markers of the present invention are highly useful for the direct selection of desirable fad2 and fad3 alleles during marker-assisted trait introgression and breeding. In a one aspect of the embodiment, two single nucleotide polymorphism (SNP) markers are provided which correspond to the alleles. Thus, the present invention advantageously permits one of skill in the art to breed for the molecular markers described herein, or derivatives thereof, rather than breeding for a high oleic / low linolenic phenotype.

The present invention relates to methods for detecting JAK2 nucleic acid in acellular bodily fluid samples from patients with neoplastic disease and determining if the nucleic acid contains one or more mutations or one mutation and one deletion. The methods are useful for diagnosing patients that have cells with mutations in the JAK2 gene that effect kinase activity. The detection of such mutations can be used to determine treatment for patients or stratifying patients for therapy and management.

The identification of a novel mutation in the testis specific Y-like gene and association of the mutation with SIDDT syndrome are disclosed. Methods for diagnosing SIDDT syndrome are disclosed. Methods for identifying compounds for use in the diagnosis and treatment of disorders associated with mutation in the TSPYL gene are also disclosed. The invention therefore provides nucleic acid sequences, genes, polypeptides, antibodies, vectors containing the gene, host cells transformed with vectors containing the gene, animal models for the disease, methods for expressing the polypeptide, genetic screening methods and kits, diagnostic methods and kits.

The invention relates to a kit for detecting genetic cardiac hypertrophy related gene mutation samples, particularly to a product for detecting genetic cardiac hypertrophy related genes by a massively parallel sequencing platform technology. The method involved in the kit comprises: a) uniquely designing and preparing a capture probe, which is directed at all exon fragments of genes ACTC1, ACTN2, BRAF, CALR3, CASQ2, CSRP3, GLA, HRAS, JPH2, KRAS, LAMP2, LDB3, MAP2K1, MYBPC3, MYH6, MYH7, MYL2, MYL3, MYLK2, MYOZ2, PRKAG2, RAF1, SOS1, TCAP, TNNC1, TNNI3, TNNT2, TPM1, TTN, TTR, and VCL; b) designing unique joints with tag sequences; c) using universal primers to conduct PCR amplification on probe sequences; and d) designing unique operations for capture of mixed target fragments. The massively parallel sequencing platform prepared by the method and the kit has the advantages of high sample throughput, high efficiency, and easy operation, thus greatly reducing the sequencing testing cost.

The invention discloses a tobacco polyphenol oxidase gene NtPPO1 and a site-directed mutagenesis method and application thereof. The nucleotide sequence of the tobacco polyphenol oxidase gene NtPPO1 is shown in SEQ ID:No.1, the coded amino acid sequence is shown in SEQ ID:No.2. The invention further discloses a clone method of the tobacco polyphenol oxidase gene NtPPO1. The clone method comprisesthe specific steps that firstly, cDNAs of tobacco leaves are synthesized; secondly, PCR amplification of the NtPPO1 gene is conducted; thirdly, a CRISPR / Cas9 carrier of the NtPPO1 gene is constructed;fourthly, sequencing detection of the NtPPO1 gene mutation is conducted. The NtPPO1 gene has a wide application prospect in preventing tobacco browning. The tobacco polyphenol oxidase gene and the encoded protein thereof provide the gene and technology support for crops especially for anti-browning breeding of tobaccos.

Genetic polymorphisms can change protein structure and function, altering dispositions to diseases and conditions. A single nucleotide polymorphism is the smallest genetic mutation and the most difficult to detect. However, single nucleotide polymorphisms also make up 90% of known genetic mutations, thus identifying such polymorphisms is essential. Single base extension uses the affinity of one base for its complementary base to detect polymorphisms, including single nucleotide polymorphisms. Planar waveguides are used as the platform for single base extension enabling rapid, real time detection of genetic polymorphisms. Detection limits in the picomolar range can be obtained. Signals from the non-matched DNA bases are in the range of the blank signal. Detection times of 5 minutes are reported.