386 results about "Multi gene" patented technology

A multigene expression vehicle (MGEV) consisting essentially of a polynucleotide comprising 2 to 8 domain segments, D, each domain encoding a functional protein, each domain being joined to the next in a linear sequence by a Linker (L) segment encoding a Linker peptide, the D and L segments all being in the same reading frame, and at least one of the domains is not a type two protease inhibitor.

The invention discloses a method for expression of CRISPR sgRNA by a eukaryotic cell III-type promoter and a use thereof. A eukaryotic cell III-type promoter (U6 or H1) starts expression of multiple hairpin structure small RNAs connected in series by Drosha cleavage sites. Through expression in the eukaryotic cell, multiple CRISPR sgRNAs with bioactivity can be produced. Aiming at the U6-sgRNA-shRNA-shRNA structure, the U6-started expression structure can produce a primary transcript containing multiple sgRNAs and shRNAs. In the eukaryotic cell, through processing, the sgRNAs can respectively identify corresponding target sites so that the Cas9 protein is guided to target multiple sites and thus a foundation is laid for multiple gene editing. Compared with the traditional method for expression multiple sgRNAs and the tRNA-gRNA system new reported by Plants, the method provided by the invention has a simple structure and is convenient for construction. Through a Golden Gate method, shRNA-sgRNA sequences are connected in series so that more sites are targeted or multiple shRNA interference genes are expressed.

The invention is an automated multiple-purpose, integrated laboratory system comprising interchangeable modular elements for the construction and measurement of biological assays. The functions of the modular elements may include multiwell platform handling, chemical reagent or cell management, volumetric transfer of liquids for assay construction or for recovery of reaction products for analysis, incubation under controlled environmental conditions, measurement of spectrometric signals originating from the assays, processing and analysis of the resulting spectrometric data, and other functions. The modular elements are arranged around a number of robotic elements that deliver plates to different modular elements, transfer plates to groups of modules served by a different robotic element, or other actions necessary in plate handling. Liquid transfer to and from multiwell platforms, necessary for assay construction or for the initiation of physiological events in cells, is partitioned among different modules specialized for transferring nanoliter or smaller volume quantities of chemical concentrates, or microliter quantities of assay reagents, cells, media and other assay constituents. Applications of this invention include the quantitation and analysis of the expression of multiple genes in cells, measurement of multi-gene expression kinetics, analysis of activation or suppression of multiple signal transduction pathways, screening chemical compounds for modulatory effects on multi-gene expression or on signal transduction pathways or on other biochemical networks of cells, or other analytical biological or biochemical assays.

The invention belongs to the field of immunotherapy and relates to a universal CAR-T cell and a preparation method and application thereof. In the universal CAR-T cell, the functions of a T cell antigen receptor (TCR) and major histocompatibility complexes (MHC I and MHC II) in the T cell are inhibited while multi-gene knockout is performed; a gene encoding the TCR includes TRAC and / or TRBC; genesencoding the major histocompatibility complexes include HLA-A, B2MH and CIITA. The universal CAR-T cell can target relevant markers of specific tumors and inactivate the functions of the TCR and theMHC on the cell surface, can reduce immunological rejection caused by allogeneic cell therapy and safely and effectively remove tumor cells in the diseased human body, is not affected by the disease or a treatment mode of a patient in use and can be prepared at any time, treatment can be provided at the optimum time, and treatment effectiveness is ensured.

The invention provides a cultivation method of Cyp gene knocked-out rats, and a preparation method of liver microsome of the rats. The Cyp gene knock-out herein includes Cyp single gene knock-out and Cyp multiple gene knock-out. In the method, firstly a Cyp gene knocked-out rat is constructed by means of a CRISPR / Cas system, which includes selection of a knocked-out target site, in-vitro synthesis and transcription of sg RNA and Cas9m RNA, preparation of a pseudopregnant female rat, in-vitro micro-injection and transplanting of a single-cell embryo, and cultivation of the rat, and finally, a homozygote Cyp gene knocked-out rat can be cultured. Furthermore, the liver of the Cyp gene knocked-out rat is extracted and is subjected to homogenization and differential centrifugation to prepare the liver microsome of the rat in Cyp gene deletion. The invention also provides an application of the Cyp gene knocked-out rats and the liver microsome thereof in study on drug metabolism.

The invention discloses a preparation method of a staphylococcus aureus CRISPR / Cas9 system and an application of the system in constructing a genetically modified mouse model. The staphylococcus aureus CRISPR / Cas9 system is composed of two components, namely Cas9 mRNA and gRNA, wherein a preparation method of the Cas9 mRNA is achieved by adding a T7 promoter to the upstream region of original Cas9 coding DNA, and a preparation method of the gRNA is achieved by adding the T7 promoter to the upstream region of an original gRNA coding sequence. The staphylococcus aureus Cas9 mRNA and gRNA, which are injected to mouse fertilized embryos through micro-injection, can achieve gene editing and modification of various types, such as single-gene knockout, multi-gene knockout and / or gene knock-in and the like, on the mouse fertilized embryos; therefore, the CRISPR / Cas9 system has a good application prospect in the aspects of fertilized embryo gene editing and modification of such animals as mouse and the like as well as construction of animal models.

A statistical and functional correlation strategy to identify changes in cellular pathways specifically linked to impaired cognitive function with aging. Analyses using the strategy identified multiple groups of genes expressed in the hippocampi of mammals, where the genes were expressed at different levels for several ages. The aging changes in expression began before mid-life. Many of the genes were involved in specific neuronal and glial pathways with previously unrecognized relationships to aging and / or cognitive decline. The processes identified by the strategy suggest a new hypothesis of brain aging in which initially decreased neuronal activity and / or oxidative metabolism trigger separate but parallel genomic cascades in neurons and glia. In neurons, the cascade results in elevations in calcium signaling and reductions of immediate early gene signaling, biosynthesis, synaptogenesis and neurite remodeling. In contrast, glia undergo increased lipid metabolism and mediate a cycle of demyelination and remyelination that induces antigen presentation, inflammation, oxidative stress and extracellular restructuring. These identified genes and the proteins they encode can be used as novel biomarkers of brain aging and as targets for developing treatment methods against age-related cognitive decline, Alzheimer's Disease and Parkinson's Disease.

Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5′ promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all non-essential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. Additional genetic elements encoding inteins and additional proteins can be fused in frame to the 5′-terminus of the coding region for the second protein to form a construct for expression of more than two proteins. A single 3′ termination sequence, such as a polyadenylation sequence when the construct is to be expressed in eucaryotic cells, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits, illustrated by glyphosate tolerant plants producing BT toxin, and / or value added products, illustrated by the production of polyhydroxyalkanoates in plants.

The invention provides a systemic detection method of a tumor marker and application thereof. The method comprises the following steps: 1) providing a sample from a checking object, wherein the checking object is a person; 2) detecting the sample of step 1), wherein in the detection step, whether various protein tumor markers exist or not is determined by utilizing high-throughput nucleic acid sequencing and determination comprising polygene mutation, polygene methylation, a plurality of micro RNAs (Ribonucleic Acid) and the like and utilizing a protein chip. According to the systemic detection method of the tumor marker and the application thereof, the different tumor markers are subjected to systemic detection including DNA (Deoxyribonucleic Acid), RNA, protein and the like, so that thesensitivity of early-stage detection of the tumors is improved.

The invention is nerve net system which can realize heredity arithmetic. It is made up of computer nerve net model component and the interfaces. The character lies in: 1. after that the computer setsthe population size, coding type and length, heredity operation probability and the arithmetic ending condition of the arithmetic, the nerve net uses population size to realize the whole heredity operation including selection, crossing, mutation and personal adaptability value, and outputs the optimized calculation and result through computer; 2. designs the heredity operation nerve net model which can realize multi-father crossing operation and multi-gene mutation operation, realizes the two operation of two-value coding heredity arithmetic and real number coding heredity arithmetic.

Polygenic disorders are due to the additive effect of multiple genes interacting with the environment. Because of the small effect size of each gene and considerable genetic heterogeneity, when single genes are examined, the outcome of association and linkage analyses are variable from study to study. Techniques are needed that take these unique characteristics of polygenic disorders into consideration. The present invention discloses that the formation of a polygenic score, consisting of the additive effect of multiple candidate genes, and its assessment using receiver operating characteristic (ROC) plots, provides such a technique. Six genes previously shown to be associated with Alzheimer's disease were examined, APOE, ACE, ACP1, ESR1, PNMT and SLC6A4. The total fraction of the variance, the area under the ROC plots, and the range of risks were similar for both groups indicating that despite genetic heterogeneity and the small effect size of most genes, consistent risk analyses could be obtained by examining the additive effect of these multiple genes. The present invention also discloses diagnostic tests for determining a subject's risk of developing Alzheimer's Disease or specifically Late Onset Alzheimer's Disease.

The invention discloses establishment and application of a plant multi-gene knockout vector. A method of the establishment of the plant multi-gene knockout vector comprises the following steps of (1) establishing a binary expression vector pC1300-Cas9; (2) establishing an intermediate vector SK-9RNA; (3) establishing single objective gRNA; and (4) serially connected a plurality of pieces of gRNA and establishing a final binary expression vector. A plurality of gRNA sequences are connected to the binary expression vector of an existing Cas9 sequence by an isocaudamer connecting method, and multiple plant mutants can be obtained by an agrobacterium tumefaciens infected transgenic technology. By an isocaudamer connecting establishment strategy, pieces of gRNA can be combined, and multiple plant mutants can be obtained by transgenosis once, so that an efficient and convenient multiple plant mutants preparing method is established.

African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal viral disease of domestic pigs. Control of ASF has been hampered by the unavailability of vaccines. Experimental vaccines have been derived from naturally occurring, cell culture-adapted, or genetically modified live attenuated ASFVs; however, these vaccines are only successful when protecting against homologous viruses. Among viral genes reported to be involved in virulence are components of the multi gene family (MGF). Here we report the construction of a recombinant ΔMGF virus derived from the highly virulent ASFV Georgia 2007 (ASFV-G) isolate. In vivo, ASFV-G ΔMGF administered intramuscularly (IM) to swine at either 102 or 104 HAD50 are completely attenuated; the inoculated animals are completely asymptomatic. Animals infected with 102 or 104 HAD50 of ASFV-G ΔMGF are protected against the presentation of clinical disease when challenged at 28 days post infection with the virulent parental strain Georgia 2007.

The invention relates to the field of genetic engineering and biotechnologic detection, and concretely relates to a gene group and a kit for diagnosing lung cancer, and a diagnosis method thereof. The gene group for diagnosing the lung cancer includes ABCB1, AKT1, ALK, APC, ATIC and other 63 genes. The method using the gene group to diagnose the lung cancer comprises the following steps: (1) extracting free DNA and genome DNA from the plasma of a sample to be detected; and (2) breaking the genome DNA into fragments with the length of 150-250 bp, carrying out hybrid capture on the broken genome DNA and the free DNA to construct a DNA library, carrying out online sequencing, and analyzing the obtained sequencing result. Exon and partial intron regions of the 68 genes of the free DNA are enriched at one time by a probe capture technology to realize multi-gene and multi-target parallel deep high-throughput sequencing with high accuracy, optimize the detection flow and improve the detection precision, so liquid biopsy, low-frequency detection and tumor diagnosis become possible.

The invention relates to a method for performing multi-gene disease inheritance risk comprehensive assessment to an individual. The method comprises the following steps that: inheritance risk factors of a special multi-gene disease in a special crowd are screened; the risk degree of each risk factor to the special multi-gene disease is determined; and resources such as NCBI, HapMap databases and so on, are utilized to determine the frequencies of gene types of polymorphic gene type loci in the special crowd. The method is characterized in that: an assessment model is established, and the risk rank of the multi-gene disease of the individual is assessed from the genetic angle. The method is a comprehensive, objective, easily-operated, and intellectualized risk assessment method.

The invention discloses a combined primer for screening human deafness gene mutation. The combined primer comprises a combined forward primer and a combined reverse primer, wherein the combined forward primer consists of two parts namely a joint part P1 sequence and a forward primer sequence, and the combined reverse primer sequentially consists of a joint part P2 sequence, a label sequence and a reverse primer sequence. The invention also discloses a kit for screening the human deafness gene mutation. The invention also discloses applications of the combined primer and the kit in preparation of a reagent for screening the human deafness multi-gene mutation. A primer containing the label (Barcode) sequence is used for differentiating different samples, and at least 32 samples can be detected by performing a method provided by the invention once, so that combined primer can be widely applied to general investigation of deafness susceptible genes. The price of the combined primer disclosed by the invention is far lower than that of imported instruments or reagents, so that the sequencing cost is greatly reduced and then the detection cost is also greatly reduced.

The invention relates to a multi-gene high rice blast resistance material breeding method adopting marker assisted selection, and belongs to the field of rice breeding. According to the method, specific molecular marker assisted selection of rice blast resistance genes pita and pi5 is adopted, field resistance identification is combined, and accordingly, a new disease-resistant variety can be bred effectively. The method comprises implementation modes as follows: 1, parents P1 and P2 containing the rice blast resistance genes pita and pi5 respectively are hybridized with a susceptible parent Liaoxing 1 to establish a resistant and susceptible offspring group; 2, extraction of DNA of single resistant and susceptible offspring, PCR (polymerase chain reaction) amplification and electrophoretic analysis of different markers are performed; 3, the parents P1 and P2, the susceptible variety Liaoxing 1 and the single plant of the hybridized offspring group are planted in a rice blast high incidence area Donggang, Dandong, and leaf blast and spike and stem blast resistance identification is performed at a seedling stage and a maturation stage respectively. In combination of the resistance gene marker selection, the new multi-gene rice blast resistance variety containing pita and pi5 is directionally improved and cultivated.

The invention discloses a method for qualitatively and quantitatively detecting nucleic acid based on a high-flux sequencing technology. The method comprises the following steps of: preparing a single-chain DNA (Deoxyribose Nucleic Acid) library with a method for capturing a specific nucleic acid target through multi-gene multi-region two-step and one-direction amplification; sequencing the obtained single-chain DNA library based on a high-flux sequencing technology; and qualitatively and quantitatively analyzing nucleic acid according to a sequencing result to obtain a qualitative and quantitative detection result of nucleic acid. Compared with an ordinary real-time Q-PCR (Quantitative Polymerase Chain Reaction) quantifying method, the method has the advantages of lower cost, wider application, increase in the quantifying sample flux of nucleic acid and capability of accurately qualifying and quantifying the series of nucleic acids.

The present invention discloses a method breeding and application of a new recessive dwarf rape. Brassica napus L variety Jinzhou rape No.2 and de nong zheng cheng company-rape No.5 are used as parents for hybridization to obtain F1, F1 is bagged for selfing breeding to obtain segregating populations. By many years of in-field phenotypic characterization of the segregating populations, a dwarf single plant with excellent comprehensive agronomic traits and quality is selected for continuous selfing breeding for many years to breed a dwarf strain with stable dwarf trait. The dwarf material is used for hybridization with a high material, multi-generation joint analysis finds that the dwarf trait of the rape are controlled by two pairs of additive-dominance-epistatic main gene and additive-dominance-epistatic polygene, and the dwarf trait is recessive. The new dwarf material is used for hybridization with the high material to breed a semi-dwarf anti-lodging new rape variety, and the yield per unit can be significantly improved.

The present invention concerns prognosis for glioblastoma and / or assessment of the response of an individual to therapy for glioblastoma treatment. In particular, expression analysis of two or more specific genes provided in the invention is determined to predict outcome for the individual and / or to predict if the individual will respond to therapy, such as chemoradiation, for example. In specific embodiments, a multigene set from a sample from the individual is compared to a reference set of housekeeping genes.

The invention discloses a detection kit for coronavirus SARS-CoV-2 and a mutant strain thereof. The detection kit comprises a digital PCR reaction solution, a digital PCR primer mixed solution and a negative control solution, and is combined with a full-automatic microdroplet chip digital PCR system to detect the coronavirus SARS-CoV-2 and the mutant strain thereof. Specific primer probes are designed for two target genes of an ORF1ab gene and an N gene of a non-mutant strain of the coronavirus SARS-CoV-2 and three mutation sites of E484K, L452R and D614G in an S gene of the mutant strain of the coronavirus SARS-CoV-2, the detection accuracy of the kit is improved through digital PCR detection of different sites of multiple genes, and rapid judgment is achieved. The kit better meets clinical requirements in the detection, screening and vaccine development of the novel coronavirus, and also enables the detection to be faster and the application to be wider.

A method for assessment of the suitability of and / or effectiveness of a target therapy for a TNF-mediated-related disorder, such as severe or persistent asthma, in a subject evaluates the presence, absence, and / or magnitude of expression of one or more genes corresponding to contacting the sample with a panel of nucleic acid segments consisting of at least a portion of at least one member from the group consisting of the nucleotide sequences corresponding to at least one of TNFRSF1A SNP rs4149581 (SEQ ID NO:1), TNFRSF1 B SNP rs3766730 (SEQ ID NO:2) or TNFRSFI B SNP rs590977 (SEQ ID NO:3) SNPs which results in a determination that one or more of said SNPs in a sample are in linkage disequilibrium (LD). The method enables identification of the effectiveness of target therapies prior to or after starting a patient on such therapies.

The invention discloses gene sets and a scoring model for evaluating a tumor microenvironment, and application of the gene sets, and belongs to the field of biological information. It is proved for the first time that the gastric cancer has remarkable tumor microenvironment typing, and prognosis difference of gastric cancer patients with different typing is remarkable. According to a constructionmethod of the multi-gene scoring model for evaluating the gastric cancer tumor microenvironment, 44 gene sets capable of representing different microenvironment types are screened out, and the prediction efficiency of the tumor microenvironment multi-gene scoring model corresponding to the gene sets is remarkably better than that of other existing models; and the model is not only a prognostic marker of a gastric cancer patient, but also has a quite remarkable prediction effect in pan-cancer. The model provided by the invention not only can be used as a good prognosis biomarker, but also can be used for predicting checkpoint inhibitor immunotherapy response of various tumors by effectively evaluating the tumor microenvironment, and has important significance and clinical application valuefor better screening immunotherapy benefiting people.

The invention discloses a method for enriching target regions of 168 genes based on multi-probes. According to the invention, 168 genes originated from a TCGA database, targets of conventional approved targeted drugs for lung cancer, targets of targeted drugs for other tumors and targets of targeted drugs in clinical experiments are combined together, and multi-target sequences of the 168 genes are enriched in one shot by using probe capture technology; so multi-gene multi-target parallel in-depth high flux sequencing is realized, a plurality of variation types like mutation, deletion, insertion, fusion and copy number amplification of genes are examined at the same time, and the method is superior to the prior art in detection depth and width.

The invention discloses a non-invasive detection method and a kit for early screening a bladder cancer by multi-gene combination. The combined genes include APC, NID2 and p16, and specific methylationprimers are designed to detect three nucleotide sequences of target gene fragment methylation of the APC, the NID2 and the p16, so that early screening and auxiliary diagnosis of the bladder cancer are achieved. Methylation levels of three genetic markers of the APC, the NID2 and the p16 in urinary sediments are analyzed by a three-channel fluorescent quantitative PCR (polymerase chain reaction)method, and a positive or negative result of a sample is judged according to a reported CT (computed tomography) value. The non-invasive detection method can detect 90% of early bladder cancers, 64% of precancerous adenomas (larger than or equal to 1 centimeter) in preclinical study, specificity reaches 100%, invasiveness is completely omitted, the method only needs to collect urine of 50mL, and acceptability of throngs wearing no symptoms is high.

Disclosed are stable recombinant multi-gene nucleic acid constructs, such as plant binary vectors, comprising (i) a gene encoding gamma-glutamylcysteine synthetase and (ii) a gene encoding glutathione synthetase, plus preferably at least one, preferably two, genes which encode enzymes involved in the redox cycling of glutathione between its reduced and its oxidised forms e.g. glutathione reductase and / or glutathione peroxidase. Preferably the promoters linked to the genes are different and of different strengths, and may optionally be inducible. Also provided are related materials and corresponding methods and uses e.g. in plants to improve oxidative stress tolerance enhance root development, or to increase the post-harvest shelf life of the plant or part thereof.

The invention discloses a preparation method and application of a humanized CD40 gene remolding animal model. The invention also provides a sgRNA sequence capable of specifically targeting the CD40 gene, a method for preparing a multi-gene humanized animal model and related applications. According to the invention, a tool mouse suitable for transplantation of a humanized cell or tissue and a preparation method of a novel humanized animal model are provided, a research of related diseases is facilitated and an effective technical means is provided for the development of biomedical experiments.The invention also relates to remolding an non-human animal by using a humanized gene, in particular to remolding a rodent by using the gene. Particularly, remolding a mouse by using the gene relatesto the preparation method of the humanized CD40 gene remolding animal model and the application thereof in the field of biomedicine.

The invention discloses a probe preparation method for multi-gene capture sequencing. The method at least comprises the following steps: step 1, preparing a probe; step 2, capturing target DNA; step 3, performing an elution program; step 4, performing post-amplification on the captured target DNA by using a high-fidelity enzyme reaction amplification system; step 5, purifying the amplification product by using magnetic beads, quantifying to 20ng / ul by using Nanodrop, split charging and preserving; and step 5, detecting a positive control sequence by using a Real-time PCR method, and verifying the capture efficiency. A single-chain DNA capture probe is synthesized by utilizing a chip technology and comprises a single-chain biotin-labeled oligonucleotide probe pool obtained by DNA chip synthesis, probe cutting, nucleic acid amplification and DNA melting.

The invention belongs to the technical field of genetic engineering, and particularly relates to a multi-gene locus simultaneous editing method based on an endogenous CRISPR-Cas system of zymomonas mobilis and application thereof. The method mainly comprises the following steps of: constructing a plasmid containing an artificial CRISPR expression unit; selecting a plurality of target editing target loci, and designing a gRNA sequence; connecting a plurality of gRNA sequences in series, and constructing the gRNA sequences on the plasmid containing the artificial CRISPR expression unit; constructing a donor DNA sequence on a vector to obtain an edited plasmid; and transforming the edited plasmid into competent cells for editing. The method constructs a gene editing platform by utilizing theendogenous CRISPR-Cas system of zymomonas mobilis, breaks the limitation of low efficiency of exogenous CRISPR-Cas9 genome editing in the strain, realizes rapid and efficient knockout of multiple genes in the strain, and promotes the development of metabolic engineering, systemic biology and synthetic biology.