1041 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.

The invention relates to the technical field of molecular markers, and discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to the litter size character of pigs and application of the SNP molecular marker. The SNP molecular marker is located at a 34977540bp site of a chromosome 2 of an international pig reference genome Sscrofa11.1 version, and the nucleotide sequence of the site is G or A and is located in an exon region of an LUZP2 gene. The invention also discloses an application of the SNP molecular marker in breeding of the litter size character of the boar, and provides a method for breeding or auxiliary breeding of the boar with high / low litter size based on the marker. The invention also provides a specific primer for identifying the SNP site. By applying the technologies, accurate prediction and efficient screening of the litter size of the large white pigs can be achieved, the breeding efficiency of excellent breeding pigs is remarkably improved, and the population improvement process is accelerated. The invention also provides important technical support for sustainable development of pig raising industry, also provides theoretical basis for research in related fields, and is expected to promote further development of pig genetic improvement work.

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 discloses a method for targeted creation of a fast-growing new tilapia strain, and belongs to the technical field of fish cultivation. The method specifically comprises the following steps: (1) taking a fragment, as shown in SEQ ID NO.1, on a third exon of a pomc gene as a gRNA target sequence; (2) designing and synthesizing a gRNA primer sequence; (3) amplifying by adopting the gRNA primer sequence and the gRNA plasmid, and then transcribing and purifying to obtain a gRNA fragment; (4) transferring the gRNA fragments into fertilized eggs of tilapia mossambica through gene editing to obtain F0-generation positive fish; and (5) hybridizing the F0-generation positive male fish with the wild-type female fish, screening out F1-generation parent fish, and culturing the F1-generation parent fish to obtain F2-generation tilapia with homozygous deletion mutation. According to the invention, the pomc homozygous knockout line is established in the Nile tilapia mossambica for the first time, the new fast-growing line is a complete gene function deletion model, the cost is lower, the efficiency is higher, and the phenotype can be stably inherited.

Disclosed are constructs, systems, and methodologies using prime editing (PE), twin prime editing (twinPE), or multi-flap prime editing to carry out site-specific and large-scale genetic modification, such as, but not limited to, insertions, deletions, inversions, replacements, and chromosomal translocations of whole or partial genes (e.g., whole gene, gene exons and / or introns, and gene regulatory regions). In certain embodiments, the disclosure provides constructs, systems, and methods using prime editing (PE), e.g., single flap or “classical” PE or twinPE or multi-flap PE, to install one or more target sites for site specific recombination in a target genomic locus (e.g., a specific gene, exon, intron, or regulatory sequence), which may then be acted on by one or more site-specific recombinases to effectuate a large-scale genetic modification, such as an insertions, deletions, inversions, replacements, and chromosomal translocations.

The invention discloses a method for constructing a Ddx58-R109C point mutation mouse model by using a CRISPR / Cas9 gene editing technology and an application of the Ddx58-R109C point mutation mouse model. The method comprises the following steps: (1) target construction: searching a CDS region of a Ddx58 gene, determining an exon part, and determining a gene mutation site; (2) obtaining a fertilized egg, and injecting Cas9mRNA, gRNA and donor oligo into the fertilized egg of the C57BL / 6J mouse together in vitro through a microinjection technology to obtain an F0-generation hybrid mouse; (3) mating the F0-generation hybrid mouse with the wild type mouse to obtain an F1-generation mouse; (4) the F1-generation heterozygous mice which are identified to be positive are subjected to mating, and homozygous Ddx58-R109C mutant type and wild type mice can be obtained by breeding obtained offspring; (5) phenotypic analysis shows that Ddx58-R109C point mutation mice can have pathological changes of systemic lupus erythematosus and lupus nephritis; the invention can provide an effective research approach and method for researchers to deeply explore the action mechanism of Ddx58-R109C mutation in diseases, and has important significance for explaining the pathogenesis of systemic lupus erythematosus and researching and developing targeted therapeutic drugs.

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 belongs to the field of biological medicines, and particularly relates to an optimized antigen peptide aiming at L858R point mutation of an EGFR exon 21 region and application of the optimized antigen peptide. Compared with a mutant peptide, the optimized antigen peptide has stronger immune cross reaction, can improve the immunoreactivity or curative effect of related tumor vaccines, and can effectively cover more patient crowds.

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 present disclosure provides mouse models comprising an endogenous Stathmin2 (Stmn2) gene that comprises a human exon 2a polynucleotide sequence.

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 discloses a preparation method of high-purity circular RNA (Ribonucleic Acid). The preparation method comprises the following steps: constructing a cyclization skeleton sequence with self-splicing activity; the cyclization skeleton sequence comprises the following modular elements from the 5'end to the 3 'end: a T7 promoter, a 5'-terminal homologous fragment, a 3 'intron-exon fragment, a 5'-internal homologous fragment, a 5 '-terminal spacer sequence, an IRES sequence, a target fragment, a 3'-terminal spacer sequence, a 3 '-internal homologous fragment, an exon fragment-5' intron, a 3 '-terminal homologous fragment and an Xba I endonuclease site; inserting the cyclized skeleton sequence into a pUC57 vector, and carrying out plasmid amplification; an Xba I incision enzyme linearization vector is utilized to obtain an in-vitro amplification template; the method comprises the following steps: carrying out an in-vitro amplification reaction by using dsRNA-free T7RNA polymerase to obtain dsRNA-free precursor RNA; cyclizing the precursor RNA to obtain crude circular RNA; treating the crude circular RNA by utilizing poly (A) or poly (U) polymerase, and adding poly (A) or poly (U) long tails to linear RNA in the crude circular RNA; and purifying by using an affinity chromatography column to obtain the high-purity circular RNA.

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.

The invention discloses application of a homozygous double V type of a body thickness associated SNP (Single Nucleotide Polymorphism) site of a belly expanding hippocampus in breeding, and belongs to the technical field of aquatic breeding. Wherein the haplotype is randomly formed by a haplotype I, a haplotype II and a haplotype III, basic groups at exons 227 and 300 of the CSF2RB gene of the haplotype I are T and C respectively, basic groups at exons 227 and 300 of the CSF2RB gene of the haplotype II are T and A respectively, basic groups at exons 227 and 300 of the CSF2RB gene of the haplotype III are G and A respectively, and basic groups at exons 227 and 300 of the CSF2RB gene of the haplotype III are G and A respectively. Compared with other haplotypes, the homozygous double V type randomly formed by the haplotype III has remarkable advantages in the aspects of average body thickness and CSF2RB gene expression, and can be used for auxiliary breeding of fast-growing new strain molecules of the belly-expanding hippocampus, so that the breeding process of fast-growing improved varieties of the hippocampus is accelerated.

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 relates to a primer of a molecular marker related to white feather traits of Shaoxing ducks and application of the primer, and belongs to the technical field of biology. A duck reference genome version of GCF015476345.1 is used, the molecular marker is located on a 5212998th basic group of a No.13 chromosome and located on an intron between a 1M exon and a No.2 exon of an MITF gene, the molecular marker has two mutations of T and C, the character is a recessive inheritable character, a T / T genotype individual phenotype is white feather, and a T / C and C / C genotype phenotype is gray feather. The molecular marker is used for screening and breeding the white-feather Shaoxing ducks, and has great economic application value and breeding value for breeding high-quality white-feather Shaoxing ducks.

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 disclosure relates to compositions, systems, and methods related to repair RNA sequences, as well as related systems for trans-splicing a target nucleic acid using a repair RNA sequence. Various compositions and systems are described herein, including compositions comprising a repair RNA sequence comprising one or more exons and / or introns, a splice donor and / or splice acceptor, wherein the repRNA is suitable for trans-splicing. The compositions, systems, and methods described herein are suitable for targeting one or more Usher syndrome-associated genes, and treating Usher syndrome.

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 porcine circular RNA, a porcine circLDHC, an overexpression vector and a construction method and application thereof, and relates to the technical field of livestock molecular biology. The porcine circular RNA is formed by cyclizing 3rd to 6th exons of an LDHC gene, and the nucleotide sequence of the porcine circular RNA is as shown in SEQ ID NO: 1. The porcine circLDHC is obtained by carrying out reverse transcription on porcine circular RNA (Ribonucleic Acid). According to the invention, a specific amplification primer is designed for circLDHC, and the real existence of circLDHC is verified through PCR (Polymerase Chain Reaction) amplification, sequencing and RNase R enzyme digestion. Fluorescent quantitative PCR analysis finds that circLDHC is highly expressed in sexual mature testis tissues of pigs, the expression level of circLDHC is related to testis maturation, and meanwhile, the expression quantity of circLDHC in testis of 75-day-old Meishan boars is remarkably higher than that of circLDHC in testis of same-day-old Duroc boars (plt; 0.01) of the substrate. The invention further finds that the over-expressed circLDHC inhibits the proliferation of the porcine sertoli cells, promotes the maturation of the porcine sertoli cells, lays a foundation for further researching the start of the puberty period of the boars, and provides a new thought for analyzing the sexual precocious puberty mechanism of the boars and improving the reproductive performance.

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.