724 results about "Exon" patented technology

Provided herein are systems and methods for analyzing biomolecules (e.g., nucleic acid molecules, proteins). A method of nucleic acid analysis can comprise: (a) providing a sample comprising a cell comprising a target polynucleotide comprising a first exon segment and a second exon segment, wherein the first exon segment and the second exon segment flank opposite ends of a splice junction site of the target polynucleotide. The method can further comprise (b) contacting the cell with: (i) a first probe, wherein the first probe hybridizes to a first target sequence of the first exon segment, and (ii) a second probe, wherein the second probe hybridizes to a second target sequence of the second exon segment. The method can further comprise (c) linking the first probe and the second probe together, thereby generating a probe-linked nucleic acid molecule comprising the first probe and the second probe. The method can further comprise (d) identifying a sequence of the probe-linked nucleic acid molecule or derivative thereof, thereby locating the splice junction site of the target polynucleotide.

Among other things, the present disclosure provides designed DMD oligonucleotides, compositions, and methods of use thereof. In some embodiments, the present disclosure provides technologies useful for repairing mutant DMD transcripts by skipping exon 51 or exon 53, so that the transcript can be translated into an internally truncated but at least partially functional Dystrophin protein variant. In some embodiments, the present disclosure provides technologies useful for modulating DMD transcript splicing. In some embodiments, provided technologies can alter splicing of a dystrophin (DMD) DMD transcript. In some embodiments, the present disclosure provides methods for treating diseases, such as muscular dystrophy, including but not limited to Duchenne muscular dystrophy, Becker's muscular dystrophy, etc.

The invention relates to the technical field of biological information, in particular to an immunoreaction evaluation method based on tumor neoantigen activity sorting. The method comprises the following steps: obtaining genome data of tumor and normal tissues through whole exon sequencing, extracting multi-dimensional features of candidate somatic mutation, and screening by combining a Gaussian mixture model and a Transform model to obtain a high-confidence mutation genome set; predicting the HLA genotype of a patient based on sequencing data, translating and mutating into a peptide fragment, predicting the binding affinity of the peptide fragment and an MHC molecule by using an XGBoost model, and calculating a new antigen activity score sequence in combination with various parameters; and finally, synthesizing a new antigen peptide fragment according to a sorting result, carrying out in-vitro co-culture to detect an IFN-gamma secretion result, and dynamically optimizing a characteristic combination coefficient through a PPO algorithm to realize intelligent iterative updating, so that the accuracy of new antigen screening and the immunoreaction prediction capability are remarkably improved.

The invention relates to the technical field of teasel breeding, and discloses a teasel germplasm resource multi-character collaborative screening breeding method, which comprises the following steps: multi-omics data acquisition: extracting teasel germplasm leaf DNA (deoxyribonucleic acid), and capturing exon regions of disease resistance and metabolism related genes by adopting a targeted sequencing technology to obtain an SNP (single nucleotide polymorphism) marker matrix; the method comprises the following steps: collecting root extracts, detecting medicinal components through ultra-high performance liquid chromatography-mass spectrometry, and constructing a metabolite abundance matrix; measuring phenotype data of plant height, root length, root weight, root rot morbidity and survival rate under drought stress; key site and metabolite correlation analysis: constructing a Bayesian network containing SNP markers, metabolite and phenotypes by using a graph database, and carrying out cross validation through a leave-one-out method. The dipsacus asper germplasm resource multi-character collaborative screening breeding method aims at solving the problem that a traditional breeding method lacks systematic integration analysis of genomes and metabolic pathways in dipsacus asper multi-character collaborative improvement.

The invention relates to the technical field of bioinformatics, in particular to a method for identifying RNA (Ribonucleic Acid) from a transposon element based on RNA sequencing data. According to the invention, the exon level of the teRNA is taken as an analysis object, compared with a teRNA site, the teRNA exon not only realizes more accurate site identification, but also retains the existing form of TE in a transcription product, including whether a splicing site and a polyadenylic acid signal are provided or not, whether the TE is chimeric with a gene or other TE sites or not and the like. And compared with the level of the teRNA transcript, as a constituent part of the transcript, the teRNA exon significantly reduces the sequence assembly difficulty of the short-read-length sequencing data. According to the method, the Bayesian model, the parametric assembly algorithm and the local site de novo assembly algorithm are combined, and through mutual correction of different assembly algorithms and repeated comparison sequencing reading length before and after assembly, the assembly false positive is reduced, so that the teRNA recognition accuracy is improved.

The invention belongs to the technical field of biology, and particularly relates to an SLC44A1 gene and / or SLC44A5 gene knockout animal model as well as a construction method and application thereof. The construction method of the animal model comprises the following steps: deleting exon regions from the fourth exon region to the thirteenth exon region of the SLC44A1 gene and / or the second exon region of the SLC44A5 gene in mouse brain oligodendroglia cells in a high-efficiency manner by utilizing gene editing and a Cre-loxP recombinase system, so as to realize the knockout of a target gene. The animal model constructed by the invention can be used for researching the regulation mechanism of the choline transporters (SLC44A1 and SLC44A5) on myelin sheath formation, deeply knowing the biological functions of the choline transporters (SLC44A1 and SLC44A5), simulating the occurrence and development processes of human related diseases, revealing the pathogenesis of the related diseases, providing a new thought for prevention and treatment of brain myelin sheath dysplasia, and providing a new foundation for the prevention and treatment of brain myelin sheath dysplasia. And an effective drug or a treatment mode aiming at the disease is screened by utilizing a gene knockout animal model, so that a new choice is provided for clinical treatment of the disease.

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 belongs to the technical field of gene engineering, and relates to a construction method of a mouse model capable of rapidly degrading MSTN protein, which comprises the following steps: sequentially inoculating a 5'homologous arm, 3xEAAAK, an FKBP1A gene synonymous mutation sequence, 3xEAAAK, a red fluorescent protein mCherry sequence and a 3 'homologous arm on a Donor vector to obtain a Donor recombinant vector; designing gRNA based on a third exon of the Mstn gene, mixing the gRNA, Cas9 protein and a Donor recombinant vector, injecting the mixture into a mouse fertilized egg, and performing in-vitro culture and transplantation to obtain an F0-generation mouse; and hybridizing the F0-generation mouse with a wild type, or continuously breeding the filial generation to obtain the knock-in mouse model. According to the invention, a dTAG targeted protein degradation technology is utilized, the obtained homozygous knock-in mouse is subjected to dTAG administration, and rapid and reversible degradation and removal of MSTN protein can be realized.

Disclosed herein includes a method of detecting RhD incompatibility during pregnancy, comprising: a) extracting DNA from a blood sample of a pregnant mother; b) performing qPCR on the DNA to determine Ct values of at least one RhD exon and at least one internal control gene; and c) calculating a ΔCt value between the at least one RhD exon and at least one internal control gene.

The invention provides gRNA (guide Ribonucleic Acid) targeting an AGT (Absorption Growth Tube) gene and application thereof, in particular, the invention provides gRNA targeting an AGT gene exon-intron splicing junction region, the invention firstly discovers that the gRNA designed by targeting the AGT gene splicing junction region (such as an intron 1 receptor, an intron 2 donor / receptor and an intron 4 donor / receptor) can effectively inhibit the expression of the AGT gene, therefore, the gRNA can be used for inhibiting the expression of the AGT gene. According to the gRNA designed by the invention, the gene editing efficiency can be remarkably improved; and / or treating a disease.

The invention relates to a double-gene rare variation and disease relevance prediction model and an establishment method and application thereof, and belongs to the technical field of biological medicines.The establishment method of the double-gene rare variation and disease relevance prediction model comprises the following steps that S1, a sample library is screened; s2, performing quality control on whole exome sequencing data (WES); s3, performing phenotype screening; s4, performing grouping design; s5, carrying out PheWAS logistic regression analysis; s6, performing Firth logistic regression analysis and verification; and S7, carrying out double-gene feature analysis and double-gene pathogenicity relevance prediction. The method for analyzing the correlation between the rare double-gene variation and all disease phenotypes is designed for the first time, and a new method is provided for screening hereditary pathogenic factors of various diseases.

The invention discloses a group of capture probes for hemophagocytic syndrome related genes, a kit, application and a library construction method. The kit manufactured by the invention can be used for detecting 33 primary hemophagocytic syndrome related genes at one time by utilizing a hybrid capture method, the probe coverage is wide, the sequencing depth is high, and the embryonic line variation of all exon regions of the designed genes can be detected. According to the invention, all related diseases of related genes of the primary hemophagocytic syndrome can be reported, and genetic modes corresponding to the diseases can be provided.

The invention discloses a method for preparing bovine MSTN gene editing cells by using CRISPR / Cas12i, and belongs to the technical field of biology. According to the invention, an sgRNA expression vector aiming at the bovine MSTN gene is constructed, and a CRISPR / Cas12i system is utilized to carry out precise gene editing on a specific target area of a first exon of the bovine primary fibroblast MSTN gene, so that mRNA of the MSTN gene cannot be normally transcribed, and MSTN gene deleted cells are obtained. Compared with other gene editing tools, the Cas12i system shows higher specificity and editing efficiency (improved by 3%-5%), and the off-target effect risk is effectively reduced. The prepared MSTN gene deleted bovine recombinant cell line is stable and efficient in editing efficiency, the cell expression level is not affected, the MSTN gene deleted bovine recombinant cell line can be widely applied to somatic cell cloning and microinjection preparation of double-muscle-hip beef cattle, and a new strategy is provided for beef cattle breeding.

The invention discloses a method for constructing an Asiatic corn borer GBP gene homozygous mutant, which comprises the following steps: amplifying and cloning an Asiatic corn borer GBP gene sequence by using RACE technology, and the nucleotide sequence of the Asiatic corn borer GBP gene is as shown in SEQ ID No.1; two sgRNA target sequences are designed on a second exon of a GBP gene sequence of the ostrinia furnacalis, target sgRNA is synthesized through in-vitro transcription, the target sgRNA and Cas9 protein are fused in vitro to form an RNP mixture, and the RNP mixture is injected into ovum of the ostrinia furnacalis; the method comprises the following steps: carrying out PCR (Polymerase Chain Reaction) amplification on a target spot by using a target spot detection primer, sequencing to confirm a mutant individual, hybridizing and selfing to obtain a GBP homozygous mutant population. The Asiatic corn borer GBP gene homozygous mutant is constructed, so that the reproduction and immune functions of the mutant are remarkably reduced, and reference and research basis are provided for later development and formation of efficient pest biopesticide.

The invention belongs to the technical field of molecular markers, and particularly relates to an SNP (Single Nucleotide Polymorphism) molecular marker of a PRRX2 gene associated with an African swine fever virus antibody level, a primer and application. The locus of the SNP molecular marker is located at the 143bp position of the third exon of the porcine PRRX2 gene with the gene version number of XM005654574.2, the SNP molecular marker has T / C polymorphism, and the nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO.3. The invention also discloses a kit for detecting the porcine PRRX2 gene with the gene version number of XM005654574.2. The molecular marker provided by the invention is closely associated with the African swine fever virus antibody level of the pig, and compared with the pig with the genotype of TC / CC at the site, the pig blood with the genotype of TT has a higher African swine fever virus antibody level; the molecular marker can be applied to breeding of high-quality breeding pigs resistant to African swine fever, so that the economic benefit of breeding pigs is effectively improved.

The invention relates to a corn ZmmtRF2a gene mutation marker, an identification primer and application of the corn ZmmtRF2a gene mutation marker in regulation and control of grain development. A natural grain mutant crk5 found in a corn field breeding line selection process is utilized, a target gene ZmmtRF2a causing crk5 grain mutation is obtained through phenotypic analysis, linkage analysis and map-based cloning technologies, the gene encodes a peptide chain release factor 2a for mitochondrial localization, the peptide chain release factor 2a affects the mitochondrial function, and the gene can be used for inducing the mutation of the crk5 grain. The development of corn embryos and endosperm and the accumulation of grain storage substances are regulated and controlled. In crk5, ZmmtRF2a lacks 5th to 7th exons, the encoded truncated protein does not have GGQ and SPF motifs, the motifs are crucial to release activity of RF, mitochondrial binding and termination codon recognition, technical support is provided for utilization of the gene, and a new way is opened up.

The invention discloses a construction method and application of a Keman ataxia rat model, and belongs to the technical field of animal models. The construction method of the Keman ataxia rat model comprises the step of mutating the 38th nucleotide of the ATCAY gene exon 3 of a rat from G to A. The constructed Kemann ataxia rat model provides a research mode tool for researching the function of the ATCAY gene in Kemann ataxia and screening or preparing medicines for preventing or treating Kemann ataxia.

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 a DGAT1 gene SNPs marker for detecting sheep milk production traits and application thereof.Nucleotide sequence variation sites of the DGAT1 gene are detected with the first-filial generation of Dongfudrisheng sheep male * Hu sheep female as an object, a general linear mixing effect model is constructed, the correlation between nucleotide sequence variation and sheep milk production traits is researched, and the DGAT1 gene SNPs marker for detecting the sheep milk production traits is obtained. And digging a molecular genetic marker for regulating and controlling the lactation character of the sheep. Three SNPs (single nucleotide polymorphisms) sites are found in total; wherein two variation sites in the first intron are named as c.191 + 411 Cgt, and the second intron is named as c.191 + 411 Cgt; t (SNP1) and c.192-440 Cgt, and c. 292-440 Cgt; t (SNP2) and a variation site in the 17th exon are named as c.1461 Cgt; t (SNP3). For the SNP1 site, the average daily milk yield of the CT genotype ewe is increased by 8.2% and 17.04% compared with that of the CC genotype ewe and that of the TT genotype ewe respectively; for the SNP3 site, the milk fat percentage of the CC genotype ewe is improved by 0.314% compared with that of the CT genotype ewe. A new molecular marker can be provided for selection and improvement of lactation traits of the milk sheep, and the molecular marker has important application prospects in sheep screening or breeding.

The invention belongs to the technical field of gene detection, and particularly relates to application of a CFTR gene mutation combination, an amplification reagent and a product in preparation of a CF risk assessment product. The CFTR gene mutation combination consists of an intron region mutation combination and an exon region mutation combination, based on the CFTR gene mutation combination, the invention further develops an amplification reagent and a CFTR gene mutation detection product, the amplification reagent and the CFTR gene mutation detection product comprise a primer combination for multiplex PCR amplification of the CFTR gene mutation combination, and the primer combination has the characteristics of high accuracy, high specificity and high sensitivity, can accurately detect related gene mutation, and provides a reliable basis for risk assessment and diagnosis of CF. When the amplification reagent and the CFTR gene mutation detection product are combined with sweat chlorine detection for use, the diagnosis rate of CF can be remarkably improved, and the amplification reagent has important application value in clinical diagnosis of CF and is expected to provide more powerful support for early diagnosis and treatment of CF patients.

The invention belongs to the technical field of genes, and particularly relates to an amplification primer group of a renal cyst disease gene PKD1, a mutation detection method and application, the amplification primer group comprises four pairs of primers for specifically amplifying first to 34 exons of the PKD1 gene; the mutation detection method comprises the following steps: acquiring genome DNA (Deoxyribose Nucleic Acid); carrying out ultra-long fragment PCR amplification on the genome DNA through the primers, carrying out electrophoresis detection, and purifying a target fragment; mixing the purified amplification product with genome DNA in proportion, constructing a sequencing library, and performing high-throughput sequencing through target area capture; and analyzing sequencing data, and screening pathogenic PKD1 gene mutation. Compared with the prior art, the PKD1 ultra-long fragment PCR amplification primer and the renal cyst disease gene PKD1 mutation detection method have the advantages that true and false genes of PKD1 can be distinguished, the diagnosis accuracy is greatly improved, the operation process is simplified, the timeliness is improved, and the detection flux is improved.

The present invention belongs to the technical field of biology. Specifically, the present invention relates to a human whole exome sequencing probe group and the use thereof.

The invention discloses a rice RGB1 gene mutant mRGB1 and application thereof, and relates to the technical field of plant genetic engineering, a gene editing technology is utilized, a rice G protein beta subunit coding gene (RGB1, LOCOs03g46650) N-terminal structural domain coding region is targeted, three and six basic groups are deleted from a first exon and a second exon respectively, the mutant is named as mRGB1, the mutation causes plant dwarfing, upright and dense ears, and the rice RGB1 gene mutant mRGB1 has the advantages of high yield and high yield. The seeds become small, and new gene resources can be provided for genetic improvement.

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.

Disclosed herein are binding moiety (e.g., antibody)-oligonucleotide (e.g., PMO) conjugates and pharmaceutical compositions that induce an alteration in an incorrectly spliced dystrophin mRNA transcript to induce DMD exon 45 skipping. Also described herein include methods for treating muscle dystrophy including Duchenne muscular dystrophy that comprises administering such conjugates or a pharmaceutical composition that induces alteration in an incorrectly spliced dystrophin mRNA transcript to induce DMD exon 45 skipping.

This vector for nucleic acid introduction contains a nucleotide sequence that encodes a Cas protein, a nucleotide sequence that encodes a guide RNA, and a nucleotide sequence of a nucleic acid to be introduced. The guide RNA targets an intron adjacent to the 5' side, with reference to a sense strand, of an exon that includes a stop codon. Also provided is a nucleic acid introduction method comprising a step for introducing a nucleic acid to be introduced through non-homologous end joining by targeting an intron that is adjacent to the 5' side, with reference to a sense strand, of an exon that includes a stop codon.

The invention relates to an SNP (Single Nucleotide Polymorphism) molecular marker related to the size of a sweet cherry fruit and application, and belongs to the technical field of genetic breeding of sweet cherries. The molecular marker disclosed by the invention is derived from an exon of FUN001609 and is positioned at a position 16366288 of a first chromosome of Prunus avium Tieton Genome v2.0, and nucleotide at the position is G or A. The molecular marker disclosed by the invention has the advantages that the molecular marker disclosed by the invention can be used for detecting the molecular marker; the SNP site is obviously related to the size of the sweet cherry fruit, and the genotype of the size of the sweet cherry fruit is AAgt; gGgt; gA. Meanwhile, a primer is designed according to the SNP site to carry out PCR amplification on the sweet cherries to be detected, and a PCR product is subjected to sequencing inspection. The SNP molecular marker disclosed by the invention can be used as an auxiliary selection marker of fruit size characters in a sweet cherry breeding process, so that the screening efficiency of large-fruit sweet cherry varieties is improved, the breeding period is shortened, and the breeding time and workload of the large-fruit varieties are greatly reduced.