293 results about "Exon" patented technology

The invention relates to the technical field of plant genetic engineering, and discloses a haplotype identification marker of a rice flood-resistant gene ARF9 and application of the haplotype identification marker. The molecular marker is derived from an exon of an ARF9 gene, is located at the 22009082bp and 22010019bp of a No.4 chromosome of rice, and has a basic group of C. The SNP molecular marker is related to the waterflooding resistance of a male parent oryza longistaminate of perennial rice, and the waterflooding resistance of the male parent oryza longistaminate with a locus genotype of CC is obviously higher than that of rice with a locus genotype of non-AA type. Therefore, the invention provides the loop-mediated isothermal amplification primer group for rapidly identifying the ARF9 gene haploid molecular marker, and the loop-mediated isothermal amplification primer group is used for identifying the ARF9 gene haploid molecular marker so as to rapidly screen out waterflooding-resistant rice. The invention provides a group of primers, the flood-resistant rice can be quickly identified by adopting the primers to perform loop isothermal reaction, the detection cost is low, the detection speed is high, the flood-resistant germplasm can be quickly screened in the rice seedling stage by adopting the method provided by the invention, a time-consuming flooding test does not need to be performed in the later stage, and the breeding process is greatly accelerated.

The invention discloses a method, a system and equipment for detecting internal tandem repeat and a storage medium, and the key points of the technical scheme are as follows: obtaining a first reference sequence according to at least one target exon sequence corresponding to a protooncogene, and obtaining a second reference sequence according to at least one target intron sequence corresponding to the protooncogene; comparing the sequencing data of the to-be-detected sample to the second reference sequence to obtain a first comparison result, extracting an uncompared sequence from the sequencing data according to the first comparison result, and comparing the uncompared sequence to the first reference sequence to obtain a second comparison result; and determining a first detection result according to the second comparison result and a first reference sequence, and performing false positive filtering on the first detection result to obtain a second detection result. According to the invention, false positive can be reduced so as to ensure the accuracy and reliability of subsequent analysis.

The invention discloses an Indel marker related to a corn kernel protein content character and application of the Indel marker, and belongs to the technical field of plant molecular genetics and crop molecular design breeding. The Indel marker is located on a gdh2 gene of a chromosome 10 of a corn genome, and insertion or deletion of a nucleotide sequence as shown in SEQ ID NO.5 occurs in an intron region between a second exon and a third exon. By applying the molecular marker-assisted selection technology, high-protein new germplasms S3317 (the crude protein content is 17.0%) and JS231148 (the crude protein content is 15.6%) have been successfully cultivated, the increase amplitudes of the high-protein new germplasms and JS231148 are both more than 15% compared with those of the conventional control, and the lysine content reaches 0.41% and reaches the national high-quality protein corn standard.

The invention provides an SNP (Single Nucleotide Polymorphism) marker for targeted identification of systemic lupus erythematosus of children and application of the SNP marker. A new SNP site (RS25671007) is identified in a child systemic lupus erythematosus patient through whole exon sequencing, the site is located at the 734th pair of basic groups of a PABPC3 gene, and mutation from C to T exists at the site, or mutation from threonine to isoleucine exists in coded amino acid. In addition, in-vitro cell model experiments prove that the SNP causes significant up-regulation of the antibody type transformation function, and the SNP can be used as a reference marker for risk stratification or medication guidance of early targeted diagnosis, detection, gene evaluation and the like of systemic lupus erythematosus of children.

Aspects of the disclosure relate to recombinant gene editing complexes comprising a recombinant gene editing protein and guide RNA (gRNA) that specifically hybridizes to a region of a C90RF72 gene (e.g., a region flanking a G4C2 repeat or within a exonic region of the gene).

The invention provides a primer probe set for digital PCR detection of non-small cell lung cancer EGFR mutation and application thereof, and relates to the technical field of digital PCR detection, the primer probe set comprises a primer pair and a probe for specific detection of EGFR gene exon 19 deletion mutation (19del) and exon 21 L858R point mutation, and the 19del mutation probe comprises a plurality of sequences to cover common subtypes. The primer probe group can be combined with a microfluidic digital PCR platform for use, a sample is divided into tens of thousands of microdroplets for amplification, and FAM, HEX, ROX and CY5 fluorescence channel signals are detected to realize absolute quantification. The system provided by the invention has high sensitivity and excellent linearity and precision, is particularly suitable for detecting extremely-low-frequency EGFR mutation in body fluid such as plasma, and has important application value in individualized diagnosis and treatment, curative effect monitoring and drug resistance evaluation of non-small cell lung cancer.

The application provides a rice heart-white gene WCG1 and application thereof, and particularly relates to positioning, cloning, function verification and application of a WCG1 gene for controlling a rice kernel heart-white phenotype. A C / A natural variation exists at a ninth exon 986 site of the WCG1 gene, the variation site makes the rice kernel present a heart-white phenotype, that is, a kernel middle heart-white and edge transparent phenotype, and the phenotype is an excellent phenotype of "sake brewing good suitable rice" specially selected in sake brewing.

The application discloses a KASP molecular marker of a Nicotiana benthamiana NbSAG101a gene and application thereof. The KASP molecular marker is an InDel insertion and deletion marker of the Nicotiana benthamiana NbSAG101a gene, is located at 256-262 bp of a second exon of the Nicotiana benthamiana NbSAG101a gene, and the nucleic acid sequence of the KASP molecular marker is shown as SEQ ID NO: 4. A wild-type allele of the Nicotiana benthamiana NbSAG101a gene is C, contains a 7 bp sequence shown as SEQ ID NO: 4, and a mutant-type allele is A, and the 7 bp sequence shown as SEQ ID NO: 4 is deleted. The KASP marker provided by the application can accurately and quickly distinguish wild-type homozygotes, heterozygotes and mutant-type homozygotes of the NbSAG101a gene, has high specificity, high accuracy, fast detection speed, low cost and high flux.

This invention discloses an efficient RNA data preprocessing method to address the problems of low processing efficiency and low information density in high-throughput sequencing data. Its core steps include: (1) introducing a parallel processing scheme for high-throughput sequence data, rapidly mapping RNA-seq data to a reference genome to generate a BAM file; (2) extracting base sequences and expression levels and storing them as compact PKL format files; (3) extracting all exon position information by parsing the genome annotation file; (4) combining multi-sample expression level data to screen for highly variable exons and constructing a high-information-density feature list based on the sample set; and (5) accurately extracting target sequences from the preprocessed file based on this list. Compared to traditional methods, this innovative approach achieves triple optimization: full-process parallel processing for accelerated computation, high-compression data storage, and adaptive feature selection. Processing speed is increased by 3-5 times, and data volume is reduced by more than 90%, making it suitable for high-throughput RNA-seq data analysis with large sample sizes.

The invention discloses construction and application of a Primrose syndrome non-human animal model, and belongs to the technical field of animal models and disease research. According to the invention, through a CRISPR / Cas9 technology, a codon CAC for coding 596th histidine in a No.14 exon of a mouse Zbtb20 gene is subjected to site-directed mutagenesis into CGC for coding arginine, and a hybrid mouse model for simulating pathogenic point mutation (p.H596R) of human Primrose syndrome is constructed. The model can stably reproduce key clinical phenotypes of the Primrose syndrome, including overgrowth after adult, serum IGF-1 rise, memory dysfunction and anxiety behaviors, and shows abnormal hippocampal neuronal development and synaptic transfer related pathways and the like. The invention provides an important experimental tool for deeply revealing the pathological mechanism of Primrose syndrome, developing drug screening and treatment intervention research and the like.

The invention provides a single-chain and double-protein-expressed circular RNA construct and a preparation method and application thereof, and the single-chain and double-protein-expressed circular RNA construct comprises 5'and 3 'introns used for realizing RNA self-splicing and cyclization as a first kind of introns; the 5'and 3 'homologous arms are used as pairing sites in RNA molecules; the exon 1 and the exon 2, which are connected after being spliced, can be spliced and connected together under the action of the first type of introns to form a complete and continuous open reading frame; an interval sequence for providing spatial and adjustment sequence structures; the IRES is used for starting translation; a glycine-serine linker as a flexible linker; a green fluorescent protein for gene expression monitoring; a luciferase for producing light by a catalytic chemical reaction. All the advantages of the double-IRES system for generating natural complete protein are reserved, and the core defect of low efficiency can be thoroughly overcome.

The invention relates to the technical field of molecular biology, in particular to a methylation marker for diagnosis of diabetic nephropathy. Specifically, the methylation marker comprises a P site of a CAT gene promoter region and / or an E site of a CAT gene exon 1 region. Based on a detection system combining methylation sensitive restriction enzyme with quantitative PCR, the kit has the advantages of simplicity and convenience in operation, good repeatability, accuracy in quantification and easiness in standardization, and is suitable for low-invasive samples such as peripheral blood. By effectively distinguishing pure type 2 diabetes patients from diabetic nephropathy combined patients, a reliable technical means is provided for risk stratification, early intervention and dynamic monitoring of diabetic people, and the method has good clinical application prospects and popularization value.

The invention relates to the technical field of molecular markers, in particular to an SNP (Single Nucleotide Polymorphism) molecular marker for predicting the daily age character of a pig for correcting 100 kg weight and application of the SNP molecular marker. The molecular marker is Ggt at the 53th site, the 157th site and the 723th site of the 12th chromosome of the Sscrofa11.1 version of a pig genome; according to the present invention, the CHD3 gene has the A single nucleotide polymorphism (SNP), and the site is located in the CHD3 gene exon region and belongs to the nonsynymous SNV; whole genome association analysis (GWAS) is carried out on a group containing 428 pigs, it is found that the site is extremely remarkably related to the growth speed required for correcting the day age of 100 kg of the weight of an individual (P = 1.27 * 10 <-10 >), and the individual carrying G allele shows higher breeding value, can serve as an important marker for molecular assisted breeding, is not limited by factors such as age and sex, and can be used as a marker for molecular assisted breeding. The method is suitable for breeding excellent boars, and can accelerate the formation of populations with excellent growth performance and improve the breeding efficiency of improved breeds.

The application belongs to the field of molecular biology, and particularly relates to a specific marker of an exon region of a Oplegnathus punctatus hmcn2 gene, a primer pair, a detection method, a kit and application. The specific marker of the exon region of the Oplegnathus punctatus hmcn2 gene is a nucleotide sequence of non-insertion and insertion specific markers of the exon region of the Oplegnathus punctatus hmcn2 gene DNA fragments, and the sequence is shown in bases of SEQ ID NO:1 and SEQ ID NO:2. The specific marker is applied to identification of female and male genetic sex of the Oplegnathus punctatus. The application establishes a method for realizing rapid genetic sex identification by rapidly detecting an exon DNA insertion variation of the Oplegnathus punctatus hmcn2 gene, and the method has a wide application prospect in the fields of sex identification of the Oplegnathus punctatus, preparation of high male seedlings and family selection breeding.

Antisense oligonucleotides for modulating UNC13A splicing (e.g., inhibiting the inclusion of UNC13A hidden exons in mature UNC13A mRNA), compositions comprising such antisense oligonucleotides, and methods of use are described. Also disclosed are pharmaceutical compositions comprising one or more antisense oligonucleotides, and methods for treating UNC13A-related diseases or diseases associated with TDP-43 dysfunction in a subject by administering the antisense oligonucleotides to that subject.

The invention relates to a full-length sequence amplification primer, an amplification method and a three-generation sequencing method of an HLA-I antigen gene, and belongs to the field of gene detection. The HLA-I antigen gene is subjected to PCR (Polymerase Chain Reaction) amplification by using 8 pairs of characteristic amplification primers. And each gene can be subjected to high-yield specific amplification on a full-length sequence containing complete intron and exon regions only through one round of amplification and enrichment, and 7.8 Kb long-fragment genes can be captured once at most. The experimental deviation caused by simultaneous amplification of a single gene by using a plurality of pairs of segmented primers and the indirect error caused by gene splicing are effectively avoided. The method comprises the following steps: carrying out quality inspection and purification on amplicon PCR (Polymerase Chain Reaction) products, mixing samples, constructing an HLA-I type antigen gene library, carrying out accurate and complete sequencing on the full-length sequence of the HLA-I type antigen gene by adopting a three-generation PacBio Sequel II platform, and ensuring that the Hifi reads base accuracy can reach 99% or above. The reading of HLA-I type antigen gene full-length sequence variation information and the haplotype analysis at a high resolution level can be realized.

Double-stranded ribonucleic acid (dsRNAi) agents that target exon 1 of the huntingtin (HTT) gene are provided.SOLUTION: A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of huntingtin (HTT), the agent comprising a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a certain specific nucleotide sequence, and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from another certain specific nucleotide sequence, provided are dsRNA agents wherein one or more lipophilic moieties are conjugated to one or more internal positions on at least one of the sense or antisense strands.SELECTED DRAWING: None

The present application relates to the technical field of gene mutation prediction, and discloses a gene mutation prediction method generated by multi-mode annotation, and the specific process comprises the following steps: carrying out mutation type annotation on input single-base mutation position information to obtain mutation basic information containing mutation types, then using an ANNOVAR annotation tool, SpliceAI splicing effect prediction software and reference mutation information of a function effect database to carry out multi-dimensional feature annotation, using Bayesian PCA based on the obtained multi-dimensional feature mutation data set to fill in the annotation data, then using an automatic engineering feature list and a separated feature selection list to carry out feature combination and screening, and obtaining a gene mutation prediction score after gradient generation tree algorithm. The present application can be used for predicting all non-synonymous exon mutations, has good performance in classifying rare benign mutations, and can identify a small amount of mutations with high pathogenic probability from a large amount of candidate mutations.

This invention relates to a primer set and method for high-throughput assessment of drug resistance levels in rodent populations. The invention provides a set of universal degenerate primers for amplifying the Vkorc1 gene in multiple rodent species. The primer set includes primers for amplifying exon 1, exon 2, and exon 3 of the Vkorc1 gene, respectively. By designing universal degenerate primers and coupling them with next-generation sequencing technology, this invention can simultaneously amplify the Vkorc1 gene coding region of at least 15 rodent species across genera and families. This significantly improves the species universality and detection throughput of rodent drug resistance monitoring, reduces the cost of large-scale screening, overcomes the limitation of Sanger sequencing in accurately analyzing heterozygotes with insertions and deletions, and enhances the ability to discover new resistance mutations. It provides a standardized and low-cost technical solution for efficient assessment of rodent drug resistance.

The invention provides a fusion transcript recognition method, device and system based on transcriptome multiple comparison data and a medium, and the core of the method is to construct a set of full-chain analysis framework oriented to multiple mapping reads to realize fusion gene detection. Comprising a pairing read segment preliminary screening module based on a loose mapping strategy, a candidate fusion construction module based on an exon graph structure, a remapping module based on an enhanced reference transcriptome, a transcript abundance estimation module based on a probability generation model, and a fusion scoring module based on fusion site specificity support degree. And a false positive inhibition module based on coverage consistency and biological filtration. According to the method, the dependence of a traditional fusion gene detection method on a unique mapping read is broken through, multiple comparative analysis on the fuzzy mapping read abandoned due to sequence homology is proposed for the first time, and high sensitivity and low false positive rate of the fuzzy mapping read are verified in simulated data and real tumor samples; the method has remarkable technical advancement, clinical applicability and expandability.

The application belongs to the technical field of animal genetic engineering, and particularly discloses a construction method of C1R gene humanized animal cells and animal models and application thereof. The construction method of the C1R gene humanized animal cells comprises the following steps: introducing a human C1R gene into non-human animal cells, so that the human C1R gene is expressed in the non-human animal cells to produce humanized C1R protein, and meanwhile, the expression of an endogenous C1ra gene in the non-human animal cells is reduced or eliminated. The application preferably adopts CRISPR / Cas9 gene editing technology to accurately insert a genomic sequence encoding a human C1R mature protein into a 2nd exon region of a mouse C1ra gene, while retaining a mouse source promoter, a 5' untranslated region (UTR) and a signal peptide sequence. A mouse cell or model capable of specifically interacting with an anti-human C1R antibody is successfully constructed.

The invention discloses application of an SNP (Single Nucleotide Polymorphism) marker influencing the lambing number of goats. The marker is located on a PIEZO1 gene on a capa circus 18 # chromosome, the specific SNP marker site is A / C base mutation at the 236bp position of SEQ ID NO: 1 in a sequence table, and the lambing number of CC genotype and AC genotype goat individuals at the site is remarkably higher than that of AA genotype individuals. According to the present invention, the Musheng black goats are adopted as the research object, the DNA sequence of the 33rd exon of the PIEZO1 gene is subjected to PCR amplification, the sequence has an SNP variation site, the influence of the site polymorphism on the goat lambing number is analyzed, the individual Musheng black goat breeding is performed according to the influence, and the method can be used for improving the Musheng black goat lambing number so as to provide the basis for the breeding of the Musheng black goats. The breeding of a new variety (strain) taking the Muscheng black goat as a breeding material is accelerated, and a marker resource is provided for marker-assisted selective breeding of goat lambing number traits.

The invention belongs to the technical field of gene editing, and particularly relates to sgRNA for TYROBP allele knockout and application of the sgRNA. According to the present invention, the action site of the sgRNA is located on the first exon of the human TYROBP gene, and specifically comprises the TYROBP-gRNA2 represented by the nucleotide sequence such as SEQ ID NO. 2; on the basis of a CRISPR / Cas9 technology, allele editing of the human TYROBP gene is achieved through specific sgRNA, and the allele knockout efficiency of the human TYROBP gene reaches 90% or above. Therefore, according to the technical scheme, the knockout efficiency of the TYROBP gene is improved, the construction cost of the TYROBP allele knockout cell strain is reduced, and the TYROBP allele knockout cell strain has actual popularization and application value.

Recombinant polynucleotides and vectors containing an engineered (artificial) exon-intron-exon gene structure in a transgene are provided, which undergoes splicing when it is expressed in a target cell.

The invention discloses a construction method of a Klf6 gene knockout mouse embryo model, which comprises the following steps: S1, preparing a gene editing solution containing Cas9 enzyme and sgRNA for targeting a third exon region and a fourth exon region of a mouse Klf6 gene by utilizing a CRISPR / Cas9 gene editing technology; s2, taking out the fertilized eggs of the mouse, transfecting the fertilized eggs of the mouse with a gene editing solution, and then performing in-vitro culture on the fertilized eggs; s3, carrying out embryo transplantation on the two-cell stage embryo obtained in the step S2 into a pregnant female mouse, and then carrying out conventional feeding; and S4, 7-8 days after embryo transplantation, taking out the uterus and stripping the embryo to obtain the Klf6 gene knockout mouse embryo model. According to the method, gene knockout is carried out by using CRISPR / Cas9 in the fertilized egg period in combination with an electrotransfection method, and compared with an existing method, the construction process time can be greatly shortened, the working efficiency is obviously improved, and the success rate is high.

The invention provides an exon copy number variation detection method, device and equipment, and relates to the technical field of high-throughput sequencing, the method comprises the following steps: inputting feature information of a to-be-detected sample in each target interval into a target hybrid expert model, and outputting a reliability score of copy number variation of the to-be-detected sample in each target interval; obtaining the reliability grade of the copy number variation of the exon interval according to the reliability grade of the copy number variation of each target interval; updating the reliability grade of the copy number variation of the exon interval of the to-be-detected sample to obtain the reliability grade of the copy number variation of the updated exon interval; and filtering the copy number variation of the exon interval, and determining an exon copy number variation detection result of the sample to be detected. According to the method provided by the embodiment of the invention, exon copy number variation can be accurately detected.

The application relates to the technical field of gene sequencing data processing and bioinformation analysis, in particular to a Web-based whole-exome sequencing tumor gene mutation detection visual analysis method and system. The system collects user sequencing data and a reference genome version through a Web interactive interface; a program is called to perform quality control cleaning and evaluation on the data, and a visual report is generated; sequence alignment is completed based on the reference genome, and a variation site is identified through algorithm iteration; biological annotation of the variation is combined with a database, a candidate pathogenic mutation set is screened out in multiple levels according to a strategy, the candidate set is projected to a visual interface, a site state is confirmed or removed in response to a manual checking instruction, and a final gene mutation detection report is generated. The application greatly simplifies the whole-exome sequencing data processing procedure, makes it easy for clinical doctors or researchers without bioinformation background to start, and improves the popularization rate and work efficiency of tumor gene detection work.

The invention belongs to the technical field of biological medicine and molecular biology, and provides a hypertrophic cardiomyopathy virulence gene MYH7c.794Cgt; the invention relates to T (p.Thr265Ile) and an application thereof. The MYH7 gene mutation is located on the ninth exon, the 794th base is mutated from C to T, namely ACC is mutated to ATC, and the 265th amino acid in the coded amino acid sequence is mutated from threonine to isoleucine. The mutation induces cardiac hypertrophy by disrupting energy metabolism-this defect occurs prior to the occurrence of systolic dysfunction. Along with increasingly prominent status of precision medicine in cardiovascular treatment, a treatment strategy aiming at an upstream pathological process (such as energy homeostasis and mitochondrial dysfunction) provides a way with a wide prospect for preventing and treating MYH7-related hypertrophic cardiomyopathy. MYH7 gene screening has important values in the aspects of promoting early diagnosis, guiding timely treatment intervention and realizing risk-based prevention and management.

Disclosed herein includes a method of determining the RhD genotype of a fetus, comprising: (a) extracting cell-free DNA from a sample of a pregnant person, wherein the extracted DNA comprises a mixture of maternal cell-free DNA and fetal cell-free DNA; (b) performing targeted multiplex amplification on the extracted DNA to amplify a plurality of target loci together in the same reaction mixture using a plurality of primer pairs, wherein at least three of the primer pairs each targets a different exon or intron of the RhD gene and the RhCE homologue gene and is each designed to amplify a target locus that comprises a variant between the RhD gene and the RhCE homologue gene; and (c) sequencing the amplicons by high-throughput sequencing and determine the RhD genotype of the fetus using the sequencing reads of the amplicons targeting the RhD gene and the RhCE homologue gene.

The invention relates to a gRNA of a targeted non-human animal Zmpste24 gene, a preparation method of a Zmpste24 gene knockout non-human animal model and a phenotype identification method. The gRNA comprises a nucleotide sequence which is complementary with a second exon region of the non-human animal Zmpste24 gene; the target sequence of the second exon region of the non-human animal Zmpste24 gene is as shown in SEQ ID NO: 4; the gRNA comprises gRNA1 as shown in SEQ ID NO: 1 and gRNA3 as shown in SEQ ID NO: 3, and the molar ratio of the gRNA1 to the gRNA3 is (1-5): (1-5). The Zmpste24 gene knockout non-human animal model prepared by the invention shows a remarkable senescence phenotype at the age of 12 weeks, and has the advantages of remarkable phenotype, good reproducibility, short experimental period and the like. The multi-dimensional phenotype identification method established by the invention can comprehensively evaluate the influence of gene knockout on the aging process.