62 results about "Gene trapping" patented technology

A novel 3' gene trap cassette is described that does not encode a marker conferring antibiotic resistance and can be used to efficiently trap and identify cellular genes. Vectors incorporating the presently 3' gene trap cassette find particular application in gene discovery, the production of transgenic cells and animals, and gene activation.

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.

The present invention describes a comprehensive system for gene discovery using retrovirus that have been engineered to exhibit increased accessibility to genomic DNA, or to mutate and identify the chromosomal target sequences of DNA binding proteins. The strategy employs the combination of retroviral integrase/DNA binding protein fusion constructs and gene-trapping methodologies. This novel technology provides the ability to establish proviral integration at any location within the genome. In addition, it allows for the generation of a collection of eukaryotic cells in which each cell contains a mutation in a target gene or sequence for a known DNA binding protein which also allow for rapid in vivo functional analysis. Sequence information obtained for genes identified using the described methods identify a collection of eukaryotic genes related by, or directly or indirectly regulated by, a given DNA binding protein.

The invention provides primers, a method and a chip for detecting alpha and/or beta-thalassemia point mutation and deletion mutation based on whole-gene capture sequencing and application of such primers, such method and such chip. The primers, the method, the chip and application thereof have the advantages that through designing of capture probes, relevant genes involved in alpha-thalassemia and beta-thalassemia are enriched and all mutation information including SNP and indel in full-length sequences of genes is detected; through addition of autosome, X-chromosome and Y-chromosome regions as well as upstream and downstream regions of coded genes as references, structure variations such as SNV and CNV are detected; compared with existing various hotspot mutation site detection technologies, the method is capable of detecting hotspot mutation information as well as some rare mutations and undiscovered new mutation types to detect and analyze full-length sequence specificity of target genes, fully covers the mutation types and makes up the defect that a conventional detection method easily causes missing detection of low-frequency mutations and rare mutations greatly.

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).

A new type of gene trap cassette, which can induce conditional mutations, relies on directional site-specific recombination systems, which can repair and re-induce gene trap mutations when activated in succession. After the gene trap cassettes are inserted into the genome of the target organism, mutations can be activated at a particular time and place in somatic cells. The gene trap cassettes also create multipurpose alleles amendable to a wide range of post-insertional modifications. Such gene trap cassettes can be used to mutationally inactivate all cellular genes temporally and/or spatially. Cells which contain the inventive gene trap cassette can be used for identification and/or isolation of genes and for the creation of transgenic organisms to study gene function at various developmental stages, including the adult, as well as for the creation of animal models of human disease useful for in vivo drug target validation.

A capture method of breast cancer related genes is provided. Probes obtained according to a construction method of a breast cancer related gene capture probe database are subjected to hybridization with a to-be-captured target genome region-containing DNA template in a to-be-detected sample. Fragments obtained by capture are subjected to high throughput sequencing; and then based on known information of breast cancer pathogenic genes, data analysis is carried out by utilizing a bioinformatics method, and thus heritable variation conditions of nucleic acid sequences in the to-be-detected sample are obtained. The provided various methods and the obtained database (or clone database) can adjust the relative capture efficiency so as to ensure consistency capture on all the target genes, and the problem that the use efficiency of an NGS sequencing platform is greatly hindered because of heterogeneity and high cost of chip capture reported in current literatures is solved.

The present invention provides methods and compositions for performing functional genomics and gene trapping using haploid cells, including haploid or hypodiploid vertebrate cells. The present invention further provides methods for identifying genes involved in cellular signaling pathways.

The invention relates to integration vectors for modifying a target genomic region comprising, in a 5′ to 3′ direction, a splice acceptor site, a 3′ hybrid recognition site, and a marker sequence (i.e., a 5′ gene trap vector); or alternatively comprising, in a 5′ to 3′ direction, a marker sequence; a 5′ hybrid recognition site; and a splice donor site (i.e., a 3′ gene trap vector). The integration vector, upon insertion into the target genomic region is capable of producing a recombinant RNA transcript that is comprised of a hybrid recognition site for a selection molecule. The hybrid recognition site of recombinant RNA produced from insertion of the 5′ gene trap vector is comprised of a 5′ hybrid recognition site derived from genomic sequence and a 3′ hybrid recognition site derived from vector sequence. The hybrid recognition site of recombinant RNA produced from insertion of the 3′ gene trap vector is comprised of a 5′ hybrid recognition site derived from vector sequence and a 3′ hybrid recognition site derived from genomic sequence. The selection molecule selects recombinant cells comprising the integration vector inserted within the target genomic region.

The invention provides a kit for detecting gene variation in a tumor FFPE (formalin-fixed paraffin-embedded) sample. The kit comprises probes for gene trapping, and the probes for gene trapping include a probe designed for an SNV (single nucleotide variant) area, a probe designed for a CNV (copy number variation) area, and a probe designed for a FUSION area; public primers include nucleotide sequence primers shown in SEQ ID NO:1; and tag primers include nucleotide sequence primers shown in SEQ ID NO:2-SEQ ID NO:5. According to the invention, in a method for detecting gene variation in the tumor FFPE sample, the genes SNV, CNV and FUSION can be stably detected at the same time.

The invention relates to a retinitis pigmentosa mutant site and an application thereof. The retinitis pigmentosa mutant site is based on clinical resources, through a gene capture chip and a deep sequencing technique, through genetics, clinical manifestation and crowd verification are used for combination, the situation that detected 35 mutant sites are retinitis pigmentosa new infective sites is successfully confirmed, a new molecular biology base is provided for disease diagnosis, and a retinitis pigmentosa diagnosis reagent kit can be developed based on the 35 mutant sites.

Novel human polynucleotides are disclosed that correspond to human gene trapped sequences, or GTSs. The disclosed GTSs are useful for gene discovery and as markers for, inter alia, gene expression analysis, identifying and mapping the coding regions of the mammalian, and particularly human, genome, forensic analysis, and determining the genetic basis of human disease.

The invention discloses a deafness-related gene capture kit and an application thereof. The invention provides a capture probe. The capture probe is as shown in 1) or 2) which are described in the specification, wherein 1) the capture probe is composed of probes as shown in sequences 1 to 113 in a sequence table; 2) the capture probe is composed of derivatives of probes in 1); and the derivative of each probe is a probe which is obtained by substituting and/or deleting and/or adding one or more nucleotides to each probe in 1) and has the same function. According to the deafness-related gene capture kit provided by the invention, target region sequencing analysis is carried out on a whole exon group or a deafness-related genome (respectively accounting for 1% and 0.01% of the whole genome)of a hereditary deafness patient; and most pathogenic mutation information of the disease is captured.

The invention provides a gene trapping kit for trapping a blood disease related gene. The gene trapping kit provided by the invention comprises probes for gene trapping, wherein the probes for gene trapping comprise probes for a target gene and probes for a reference gene. According to the invention, in CNV (copy number variation) detection based on gene trapping and sequencing, a favorable detection result can still be obtained even under the condition that no background database is used in late data processing.

The invention provides nucleotide sequences for amplification testing of HER-2 gene copy number, a kit and application thereof. The nucleotide sequences contain 8 pairs of PCR amplification primers for amplification of BRCA1 (Gene ID: 672) full-length gene and 8 pairs PCR amplification primers for amplification of BRCA2 (Gene ID: 675) full-length gene. DNA trapped by amplification can be used forsubsequent high-throughput sequencing analysis. In comparison with a probe targeting trapping technology and a multiplex PCR technology, the method of the invention is simple to operate, is low-cost,has good sequencing data uniformity, can reach more than 99% coverage with small sequencing data amount, and is helpful for providing gene detection technical support for risk assessment or medicationof genetic tumors related with breast cancer susceptibility genes and establishment of a database.

The invention provides a sequencing method based on a gene capture technology. The sequencing method has the principle that a recombinase compound is used for assisting a single-stranded DNA probe with a closed 3' end to efficiently and specifically hybridize into a double-stranded target DNA molecule under the condition of 37 DEG C, and then an affinity label on the single-stranded DNA probe is used for carrying out specific enrichment on the target DNA. The method not only has the advantages of relatively strong long probe pairing, low probe design requirement and the like of a probe hybridization capture technology, but also has the advantages of simplicity in operation, short time consumption, no need of thermal denaturation of DNA, high capture efficiency, low cost and the like of a CRISPR/dCas9 system, and the probe does not participate in subsequent amplification, so that the identification accuracy is improved. Therefore, compared with other capture technologies, the RATE-seq has the obvious advantages of simplicity in operation, high capture efficiency, extremely short consumed time (30 minutes), good stability, low cost and the like, and is very suitable for gene capture and automatic application. In addition, the RATE-seq can effectively separate and remove human host DNA in a pathological sample, and the detection rate of pathogenic microorganisms is increased.

A method of preparing gene trapping libraries, and gene targeted cells for conditional inactivation of genes, is provided. The invention provides a vector having a mutational element cassette and a gene trap cassette, each cassette having site-specific recombination sequences.

The present application provides a vector for trapping an unknown gene of Drosophila melanogaster, which is a recombinant plasmid comprising the following nucleotide sequences in this order: an artificial consensus splicing acceptor site; a synthetic “stop/start” sequence; a reporter gene; a drug resistance gene; a gene responsible for a detectable phenotype of the Drosophila melanogaster; and a synthetic splicing donor site. The present application also provide a method for trapping an unknown gene of Drosophila melanogaster by using the vector.

The invention discloses a biosensor and a construction method of a gene trapping system, and belongs to the technical field of biotechnologies. The biosensor is a transformant or a recombinant constructed by using Acinetobacter baylyi ADP1 as a chassis organism. The construction method of the gene trapping system comprises the steps that gene fragments are trapped; and the trapped gene fragments are sequenced to obtain the sequence of the trapped gene fragments. By adopting the biosensor and the construction method of the gene trapping system, a technology developed on the basis of related technologies of the prior micro genomics, based on the whole-cell biosensor and used for trapping functional genes from environmental samples is provided.

The invention discloses a probe group and kit for detecting methylmalonic academia related pathogenic genes. The invention provides a capture probe for capturing genes related to methylmalonic academia. The capture probe provided by the invention is composed of probes as shown in sequences 1-99 in a sequence table. The kit disclosed by the invention can be used for simultaneously detecting pathogenic genes of methylmalonic academia, such as MUT, MMAA, MMAB, MMADHC and MMACHC; and moreover, the kit can be used for detecting various point mutation types of methylmalonic academia. Detection of sequences of multiple genes such as MUT, MMAA, MMAB, MMADHC and MMACHC facilitates discovery of new pathogenic mutations, and a new discovery means is provided for genetic basis of methylmalonic academia.

The invention provides a probe library and a kit for detecting lung cancer related 41 genes and an application of the probe library and the kit. Wherein the probe library comprises a lung cancer related 41 gene capture probe; the number of the lung cancer related 41 gene capture probes is 1464, and the nucleotide sequences of the lung cancer related 41 gene capture probes are shown as SEQ ID NO.1-1464. When the probe library is applied to the next-generation sequencing technology, the lung cancer related 41 genes can be detected at the same time in a high-throughput mode, and the lung cancer related 41 gene capture probes have the advantages of being high in coverage rate, high in accuracy and the like. The 41 genes related to lung cancer treatment sensitivity, resistance and toxic and side effects are incorporated into detection genes of the probe library; the detection result can directly evaluate the lung cancer treatment effect and toxic reaction in advance, can help to screen outproper drugs and treatment schemes, is helpful for doctors to make treatment decisions and reasonably take drugs for patients, and can also provide basic mutation spectrum data and detection strategies for future drug development and improvement of treatment accuracy.

The invention relates to a deafness gene capture kit, which includes an L-shaped support, on which a kit is arranged, and the kit includes a box body and a box cover, and the box body includes a bottom box and a bottom box that can pass through the lower part. It is a sleeve box that is provided with a sleeve portion and is set in the bottom box and can move up and down along the vertical portion of the support member and can rotate around the vertical portion. The sleeve between the upper end of the sleeve box and the lower end of the cover The vertical part of the box cover and the support can be connected up and down and rotated around the vertical part; the bottom box is provided with elastic blocks, and the elastic blocks are preset with various diameters. Reagent bottle jack, the box is provided with a vertical partition, and a foam block is arranged in an area separated by the vertical partition, and reagent bottle placement holes of different specifications are preset in the foam block, and the vertical partition A sponge pad is provided in the area on the other side separated by the partition. The present invention prevents the kit from tipping over through the use of supports.