25 results about "Specific mutation" patented technology

A new technique for improving the secretion production of a heterologous protein by a coryneform bacterium is developed, and thus a method for the secretion production of a heterologous protein is provided. A coryneform bacterium having a secretion production ability of a heterologous protein and modified to maintain a phoS gene having a specific mutation is cultured to produce a heterologous protein by secretion.

This disclosure relates to a technique for identifying tumor-specific mutations through integrated analysis of next-generation sequencing data using machine learning models. In certain embodiments, a computer implementation method is provided, comprising: generating sequence reads from one or more samples collected from the same patient; generating variant calling files by analyzing the sequence reads corresponding to each of the one or more samples; generating a list of candidate somatic variants by comparing the variant calling files; generating a score for each of the candidate somatic variants in the list of candidate somatic variants using a classification machine learning model, wherein the score is generated based on a plurality of classifications generated by the classification machine learning model; determining the ctDNA status for the patient based on the score, wherein the ctDNA status is either positive or negative; and generating a report providing the ctDNA status for the patient.

This invention discloses a sequencing technique for enriching mutations after library construction. The co-priming amplification (Co-PCR) technique of this invention couples a mutation probe to a short P5 / P7 primer. This short primer has a low melting temperature (Tm) and cannot hybridize and amplify on its own at the reaction temperature. Only when the mutation probe hybridizes to the mutation site can it assist the coupled short P5 / P7 primer in hybridizing to the adjacent P5 / P7 sequence at the end of the pre-library, initiating amplification. Therefore, this Co-PCR technique not only amplifies a specific mutation in a specified gene but also amplifies from both ends of the original pre-library, replicating the complete sequence information of the pre-library, including sequencing adapters, indexes, and sequencing primers. The amplified products can be directly sequenced after purification. By optimizing the co-primers, Co-PCR enrichment amplification of five sites in the pre-library was achieved, resulting in nearly 100-fold enrichment.

The invention discloses a construction method and application of a basic group editor of industrial bacteria of cephalosporium acremonium based on CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-associated protein 9), and belongs to the field of gene engineering. The cephalosporium acremonium industrial bacterium base editor provided by the invention can be used for specific mutation of a specific site of the DNA level of the cephalosporium acremonium industrial bacterium. The cytosine base editor CBE constructed by the invention is a plasmid comprising a fusion protein A and an sgRNA expression element. The base editor CBE can successfully mediate C.G-T.A, is suitable for the base editor of the high-yield CPC cephalosporium acremonium industrial bacteria, utilizes the protoplast, is high in conversion efficiency, can mediate specific site mutation in the cephalosporium acremonium industrial bacteria, and has application value.

A method of treating a subject having been diagnosed as having cancer with an immunotherapy is described. The method comprises targeting one or more clonal neoantigens selected using a method comprising determining whether a tumour-specific mutation is likely to be clonal in a subject. The method comprises providing sequence data from one or more samples from the subject comprising tumour genetic material, the sequence data comprising for each of the one or more samples, and determining the likelihood that the tumour-specific mutation is clonal as a posterior probability depending on: a prior probability of the mutation being clonal, and the probabilities of observing the sequence data if the tumour-specific mutation is (i) clonal and (ii) non-clonal, in view of a tumour fraction for each of the one or more samples and one or more candidate joint genotypes.

This invention discloses the application of the VDAC2 gene or VDAC2 protein as a target in screening drugs for the treatment of retinitis pigmentosa, relating to the field of biomedical technology. The drug is one that reduces the expression level of the VDAC2 gene or inhibits the expression level or activity of the VDAC2 protein; the CDS sequence of the VDAC2 gene is shown in SEQ ID NO.1. This invention uses a rod cell-specific promoter to inhibit VDAC2 expression, which can significantly improve the survival rate of local photoreceptor cells. This method intervenes in the early stages of the disease, conforming to the natural course of RP. It strengthens the function of rod cells before a large number of photoreceptor cells die, delaying their degenerative process and indirectly protecting cone cells, thereby prolonging the patient's effective visual retention time. This overcomes the shortcomings of existing gene therapies, which are mainly applicable to late-stage or specific mutation types, and provides a therapeutic basis for retinitis pigmentosa.

The invention belongs to the technical field of agricultural biology, and particularly discloses a grape germplasm innovation method based on X-ray gradient mutagenesis and an LD50 precise prediction model, and the method comprises the following steps: selecting an annual grape branch for vernalization treatment, and trimming the annual grape branch into a single-bud stem segment; setting an irradiation time gradient, accurately calculating an LD50 value through a cubic spline interpolation model, and carrying out X-ray irradiation on the single-bud stem segment according to the LD50 value; after irradiation, performing cutting and liquid paraffin sealing, and then performing cuttage, potting and field planting treatment; performing secondary vernalization on mutagenesis plant branches, trimming the mutagenesis plant branches into single-bud stem segments, collecting tender leaves after cutting survival, extracting genome DNA, performing whole genome sequencing, and accurately distinguishing mutagenesis specific variation and natural variation. By establishing a complete technical system of standardized vernalization, gradient irradiation, LD50 accurate calculation and whole genome verification, the problems of inaccurate irradiation parameters, nonstandard material treatment, low mutation screening accuracy and the like in the prior art are solved.

The application provides a pyroglutamic acid transaminase mutant and application thereof, the mutant is obtained by mutation on the basis of wild-type pyroglutamic acid transaminase, and specific mutation includes at least one of the following sites: (1) the 13th isoleucine is mutated into arginine; (2) the 35th valine is mutated into arginine; (3) the 183rd asparagine is mutated into aspartic acid; and the mutant is suitable for industrial enzymatic production of L-aspartic acid and the process of amino acids taking L-aspartic acid as a precursor; especially, the I13R / V35R / N183D mutant obtained by modification has high positive catalytic activity and strict direction selectivity, and completely loses the reverse reaction activity of decomposing L-aspartic acid; the mutant is overexpressed with phosphoenolpyruvate carboxylase in L-threonine production strain THRS-6, and a high-efficiency L-threonine synthesis metabolic pathway is constructed, and the yield of L-threonine is significantly improved when glucose is used as a substrate.

The application discloses a kind of identification Pinb The primer sequence for amplifying the dCAPS molecular marker is shown in SEQ ID No.10-SEQ ID No.11.The application uses a strong wheat variety as a transgenic receptor, edits the variety by gene editing Pinb Gene to make it produce specific mutations and lose function, and creates a new wheat germplasm with greatly improved grain hardness and improved flour quality. Pinb The effect is stronger than that of reported single knockout of the gene, and the resistance to wheat scab does not decrease after knockout, and the content of DON toxin in flour is actually reduced.

This invention relates to the fields of molecular biology and human genetic disease detection; specifically, it relates to a reference standard for methylmalonic acidemia and its application. The reference standard comprises positive reference cells with MMACHC gene mutation sites. The genome of the positive reference cells contains one or more combinations of specific mutation sites of the following MMACHC genes: c.609G>A, c.567insT, c.658_660delAAG, c.482G>A, c.1A>G, c.80A>G, c.217C>T, c.315C>G, and c.394C>T. This reference standard provides reliable technical support for the accurate detection of methylmalonic acidemia, especially MMACHC gene-related mutations, and is of great significance in improving detection accuracy and promoting detection standardization.

This invention discloses a radiotracer targeting pan-KRAS mutant proteins, its preparation method, and its applications. The tracer uses small molecule ligands that specifically recognize multiple KRAS mutant proteins as targeting modules, and is coupled to diagnostic radionuclides through an optimized linkage system. This tracer is independent of specific mutation sites and can broadly and specifically bind to multiple high-frequency KRAS mutant subtypes, including G12C, G12D, and G12V. Based on PET / SPECT imaging, this tracer enables rapid, non-invasive, systemic, visualized, and quantitative assessment of KRAS mutant protein expression load in living tumors, overcoming the invasiveness, spatiotemporal limitations, and tumor heterogeneity challenges of traditional biopsies. It provides a novel molecular imaging tool for accurate companion diagnosis of KRAS-mutant tumors, screening of patients benefiting from pan-KRAS inhibitors, and monitoring efficacy, possessing significant clinical translational value.

Disclosed herein is a botulinum neurotoxin (BoNT) polypeptide with a modified receptor binding domain (HC) having one or more amino acid mutations that modify the binding of the BoNT to the receptor. Specific mutations and combinations of mutations are also disclosed. Isolated modified HC, polypeptides comprising the modified HC, chimeric molecules, pharmaceutical compositions, and methods of making and using the same are also disclosed. Methods of identifying additional such modified receptor binding domains, are further disclosed.

The invention provides application of K46 lactic acid of AARS2 mediated ULK1 as a target spot in preparation of an anti-kidney cancer product, so that the K46 lactic acid of the AARS2 mediated ULK1 as the target spot can be used for preparing the anti-kidney cancer product. The use comprises partial or complete knockout of AARS2 and / or ULK1 genes; the AARS2 protein activity is inhibited; the K46 gene of the ULK1 is subjected to specific mutation; the invention relates to a cell penetrating peptide aiming at a ULK1 K46 site and other methods. And inhibiting the expression of AARS2 and / or controlling the K46 lactylation level of ULK1 to play a role in resisting kidney cancer.

A recombinant antigen against Zika / Dengue virus, and a vaccine composition thereof and its use. The recombinant antigen is obtained by introducing a specific mutation into the FL fusion loop region and optionally the non-FL fusion loop region of the E protein of a wild-type Zika / Dengue virus. By introducing the specific mutation, the FL epitope of the recombinant antigen is disrupted, which can prevent the induction of antibodies against the FL epitope; thus, the purpose of reducing or eliminating the ADE effect is achieved. In addition, the recombinant antigen has the correct conformation of the E protein, has a protein expression level similar to or higher than that of a wild-type, and can induce a relatively high titer of specific binding antibodies and neutralizing antibodies in the animal body, so that effective immune protection can be provided, and good clinical application value and industrialization prospects are achieved.

Disclosed herein in one aspect is a pharmaceutical composition comprising a plurality of neoantigen peptides and a pharmaceutically acceptable carrier, each neoantigen peptide comprising a tumor-specific neoepitope capable of binding to an HLA protein in a subject, each tumor-specific neoepitope comprising a tumor-specific mutation present in a tumor, wherein (a) the composition comprises neoantigen peptides comprising tumor-specific mutations present in at least 1% of subjects in a population of subjects having cancer; (b) the composition comprises neoantigen peptides comprising tumor-specific neoepitopes that bind to an HLA protein present in at least 5% of subjects in the population; and (c) the composition comprises at least one neoantigen peptide capable of eliciting an immune response against a tumor present in at least 5% of subjects in the population of subjects having cancer.

The application provides application of a TMEM121 protein mutant in treatment of liver cancer. The application obtains a TMEM121 protein mutant through a large amount of research, wherein the TMEM121 protein mutant is that amino acids at positions 279-281 of a wild-type TMEM121 protein are mutated from LSL to AEA. The inventors find through research that the TMEM121 protein mutant with the specific mutation at the specific position has a good inhibitory effect on growth, migration and invasion of liver cancer cells, and is obviously superior to other TMEM121 protein mutants. Therefore, the TMEM121 protein mutant can be used as a potential effective drug for treating liver cancer, and provides a new thought and selection for clinical treatment of liver cancer.

The invention relates to the technical field of biology, discloses a method for improving the yield of fluorouracil through site-specific mutagenesis and application, and provides mutant cytosine deaminase containing a specific mutation site by modifying cytosine deaminase through a gene recombination technology and a site-specific mutagenesis technology. And a recombinant bacterium capable of expressing the enzyme in a host cell is constructed. The mutant enzyme or recombinant bacteria are used as a catalyst, 5-flucytosine is used as a substrate, catalytic reaction is performed under mild reaction conditions, and 5-fluorouracil is synthesized. The invention solves the problems of low flucytosine conversion efficiency, complex process flow and the like in the prior art, the catalytic efficiency of the mutant enzyme is improved, the enzyme consumption and the reaction time are reduced, and the production cost is effectively reduced. The method disclosed by the invention is efficient and economical, and the obtained mutant enzyme not only can be used for producing 5-fluorouracil in the field of medicines, but also has the potential of being applied to the fields of biosensors, environmental monitoring and the like.

The present invention provides a circular DNA vector that can be used to introduce specific mutations in a target region of a host cell. The present invention further provides methods of using the circular DNA vector to generate engineered host cells. The circular DNA vector and methods can be used to study gene function and to generate cells that produce recombinant gene products.

This invention relates to compositions and methods of treating cancer associated with mutant RAS. In certain aspects, the invention relates to antigenic RAS peptide fragments and T-cell receptors that bind to specific mutant RAS peptide fragments in the context of specific HLA types.

The application discloses a novel Fc effector-eliminated scaffold flow antibody, and belongs to the technical field of biological medicine. The antibody comprises a L234A, L235A, P238S and D265A mutation combination in a heavy chain CH2 region, and can further comprise S267E and L328F mutations, so that the Fc receptor binding capacity is effectively reduced. The antibody type can be selected from a scFv-Fc, (scFv)2-Fc, scFv / scFv-Fc and Fab / scFv-Fc and the like. The application further provides a nucleic acid sequence for coding the antibody and a preparation method, which comprises introducing specific mutations in a CH2 region of recombinant rabbit IgG, expressing in a mammalian expression system and purifying to obtain the antibody. When the antibody is applied to a flow cytometry detection reagent, the non-specific background signal caused by Fc receptor binding can be effectively eliminated, and the specificity and accuracy of detection are improved.

The application provides an ilvC mutant, a valine-producing strain and application thereof, and in the construction process, first, Corynebacterium glutamicum ATCC 13032 is used as a mutagenesis starting point, key positive mutation sites on an ilvC gene are identified from a high-valine-producing Corynebacterium glutamicum mutant strain screened through ARTP mutagenesis, and the specific mutations are as follows: arginine at the 49th position is mutated into lysine, serine at the 52nd position is mutated into threonine, glutamine at the 87th position is mutated into histidine, and glycine at the 133rd position is mutated into cysteine; subsequently, the gene editing technology is used to integrate the mutant ilvC gene fragment into the genome of Escherichia coli W3110 as a chassis strain, and the valine synthesis capacity can still be effectively enhanced; the strain effectively improves the valine production capacity by introducing the exogenous positive mutation site.

Disclosed are methods of isolating T cells having antigenic specificity for a mutated amino acid sequence encoded by a cancer-specific mutation, the method comprising: identifying one or more genes in a nucleic acid of a patient cancer cell, each gene containing a cancer-specific mutation encoding a mutated amino acid sequence; inducing autologous APCs of the patient to present the mutated amino acid sequence; co-culturing autologous T cells of the patient with the autologous APCs presenting the mutated amino acid sequence; and selecting the autologous T cells. Also disclosed are related methods of making a cell population, a plurality of cell populations, a pharmaceutical composition, and a method of treating or preventing cancer.

The invention discloses a mycobacterium tuberculosis mutant detection crRNA and application of the mycobacterium tuberculosis mutant detection crRNA. One, two or more mismatched basic groups are arranged at the 5'end of crRNA, a section of basic group which can not be completely matched with a target section is arranged at the 3 'end of crRNA, and a loop-shaped bubble structure is formed, so that the effect of distinguishing specific mutant type and wild type mycobacterium tuberculosis is achieved, and meanwhile, the mutation point of a drug-resistant gene can be determined; the method has the advantages of low detection cost, short detection period, high sensitivity and good specificity.

The application belongs to the field of genetic engineering, and relates to a construction method of a Dhx16 mutant mouse model and application of the Dhx16 mutant mouse model in disease research. The application is specifically based on a clinical patient-specific mutation c.1360C>G (p.Arg454Gly), a gRNA-donor oligo RNP complex is microinjected into a C57BL / 6J mouse fertilized ovum through CRISPR / Cas9 technology, and a Dhx16 R457G point mutation model is constructed in the mouse. The application simulates the phenotype of a human DHX16 mutation-related disease, and for the first time, reveals the correlation between the gene mutation and male sterility, provides a reliable in-vivo experimental tool for analyzing the physiological function of the DHX16 gene, elucidating the molecular mechanism of abnormal splicing function caused by the mutation and multi-system disease, and lays an important foundation for the development of gene therapy strategies for the disease.

The present invention relates to a guide RNA targeting a specific mutation in an RS1 gene, a base editor comprising same, and use thereof and, specifically, to: a guide RNA targeting a c.422G>A (p.R141H) mutation found in the RS1 gene of patients with retinoschisis; a base editor comprising same; and use thereof.