86 results about "Targeted Mutation" patented technology

The invention discloses a conjugational transfer-assisted orthogonal targeted mutation evolution system, which comprises a donor strain and a receptor strain, a donor strain intracellular muter performs targeted mutation on a target sequence to obtain a mutation sequence, a non-self-transferable conjugational transfer functional plasmid is introduced into the donor strain, and a non-self-transferable conjugational transfer functional plasmid is introduced into the receptor strain. The mutant sequence is unidirectionally and horizontally transferred from a donor strain cell to a receptor strain cell, and the complete orthogonality of the mutants in the initial donor strain and the mutant sequence in the receptor strain is realized through the orthogonality between the donor strain and the receptor strain. The results of a sacB-sucrose reverse screening experiment show that the mutation frequency of an ortho-MutaT7 system reaches 2.96 * 10 <-2 > at 96 hours, which is 1840.9 times of the natural mutation frequency of the sacB in donor cells, and the ortho-MutaT7 system generates mutation of 12 basic group change types in a target sequence. The Ortho-MutaT7 system developed by the invention realizes mutation diversity and also has orthogonality, leakage mutation and off-target mutation of mutants in host cells are fully avoided, and a direct relationship between a mutation sequence and a phenotype is established.

The invention provides an RNP compound, an electrotransfection composition and a pharmaceutical composition thereof, a CD34 + cell gene editing method using the composition, and a method for obtaining HBG gene target mutation in a CD34 + cell genome.

The presently disclosed subject matter provides for methods and compositions for treating cancer (e.g., breast cancer). It relates to mutant PIK3CA-targeted TCRs that specifically target a mutant PIK3CA peptide (e.g., a human mutant PIK3CA peptide), and immunoresponsive cells comprising such TCRs. The presently disclosed mutant PIK3CA peptide-specific TCRs have enhanced immune-activating properties, including anti-tumor activity.

The presently disclosed subject matter provides for methods and compositions for treating cancer (e.g., breast cancer). It relates to mutant PIK3CA-targeted TCRs that specifically target a mutant PIK3CA peptide (e.g., a human mutant PIK3CA peptide), and immunoresponsive cells comprising such TCRs. The presently disclosed mutant PIK3CA peptide-specific TCRs have enhanced immune-activating properties, including anti-tumor activity.

The invention relates to an erythrocyte specific antibody screening test kit expressing a single antigen from a target erythrocyte (RBC) antigen (RhD and RhCE as 2 different antigens derived from RBC and 45 or more antigens in total including some variants of these two antigens) and a preparation method of this test kit. Said antigens are transgenically produced and provide the clearest results compared to traditional methods of antibody screening. Field-targeted mutation process is applied during the preparation of the erythrocyte-specific antibody screening test kit subject to the invention.

The invention discloses a CRISPR-Cas13a (clustered regularly interspaced short palindromic repeats-CRISPR-Cas13a) double crRNA (ribonucleic acid) system which is established by using ERASE test paper and is used for detecting the rifampicin resistance of mycobacterium tuberculosis on site. The CRISPR-Cas13a double crRNA-system comprises a Cas13a protein and crRNA, or a complex formed by the Cas13a protein and the crRNA; the crRNA comprises an anchoring sequence and a guide sequence; the anchoring sequence is used for being combined with Cas13a protein; the guide sequence is used for targeting mutation target sequences of sites 531, 526, 516 and 513 in a mycobacterium tuberculosis drug-resistant gene rpoB gene; the target sequence of the mycobacterium tuberculosis is located at the 2317th site to the 2597th site of the rifampicin drug-resistant gene rpoB. On the basis of a CRSPR-Cas13a nucleic acid detection technology, result interpretation is performed through RAA amplification in combination with a line elimination method (ERASE) lateral flow chromatography test paper, and experiments prove that the kit can target high-frequency mutation sites of four rifampicin drug-resistant genotypes at the same time, has good detection sensitivity and specificity, and can be used for detecting the rifampicin drug-resistant genotypes of the rifampicin drug-resistant genotypes of the rifampicin drug-resistant genotypes of the rifampicin drug-resistant genotypes. The method can realize rapid, efficient, simple and convenient detection of mycobacterium tuberculosis rifampicin drug-resistant nucleic acid, has a sensitivity of 10 copies / [mu] L, and has an important application value.

The invention discloses an organ-like drug resistance prediction method and device and a storage medium, and relates to the technical field of bioinformatics, and the method comprises the steps: S1, constructing an initial prediction model; s2, acquiring a training data set; s3, training an optimization prediction model; s4, acquiring related data of a to-be-predicted drug; s5, obtaining a prediction result of the drug resistance of the to-be-predicted drug; according to the method, the drug resistance of the organoid to the drug can be predicted more accurately. By fusing multi-dimensional features of gene mutation, target mutation and drug functional groups, the understanding and learning ability of the model to the drug resistance mechanism is enhanced, and the reliability of drug resistance prediction is improved. The method not only can accurately capture key information of influence of gene and target mutation on drug resistance, but also can highlight unique mutant genes of the organoid on drug resistance by comparing feature differences of the organoid and a common cell line, so that an efficient and accurate calculation framework is provided for drug resistance detection.

The present disclosure provides methods and systems for targeting cell with mutations.

This invention provides a testing method, device, and storage medium for stateful network protocols. First, it accurately identifies new state transition events based on the response feedback of the server under test. Then, it extracts corresponding client request message pairs from protocol interaction messages, and further segments them according to field location rules to obtain semantically independent field units. It then identifies differences between field units with the same name to generate candidate field groups, and filters out the candidate field set that truly triggers the state transition through testing. Finally, it performs structure-aware targeted mutations on the test seed based on the candidate field set to generate test cases. By accurately locating the core fields that trigger state transitions, it solves the technical problem of insufficient granularity in trigger condition analysis, making subsequent mutations in the testing process more targeted and effectively improving the accuracy and efficiency of state exploration in fuzzy testing of stateful protocols.

The presently disclosed subject matter provides for methods and compositions for treating cancer (e.g., breast cancer). It relates to mutant PIK3CA-targeted TCRs that specifically target a mutant PIK3CA peptide (e.g., a human mutant PIK3CA peptide), and immunoresponsive cells comprising such TCRs. The presently disclosed mutant PIK3CA peptide-specific TCRs have enhanced immune-activating properties, including anti-tumor activity.

The invention discloses a machine learning-based inhibitor drug screening method for targeting KRAS G12D mutation, and the method is used for predicting the binding capacity of candidate small molecules and KRAS G12D protein targets by constructing an integrated graph neural network (GNN) and molecular feature embedded deep learning model. Compared with a traditional virtual screening method, the method has remarkable advantages in the aspects of improving the recognition capacity of KRAS G12D mutation specific small molecules and reducing the false positive rate and has high application potential and industrial transformation value, and molecular dynamics simulation results of screened compounds show that compared with existing KRAS G12D mutation targeted drugs in research, the method has the advantages that the application potential and industrial transformation value of the screened compounds are greatly improved, and the application prospect of the KRAS G12D mutation targeted drugs is widened. The screened compound has a more stable binding trend with targeting protein in MD simulation, is expected to become a new KRAS G12D targeting inhibitor, and provides new possibility for treating pancreatic cancer.

The present invention provides a TCR targeting a KIT D816Y mutation-derived neoantigen, a nucleic acid molecule comprising a nucleotide sequence encoding the TCR or a complementary sequence thereof, a vector containing the nucleic acid molecule, and a cell transduced with the nucleic acid molecule or the vector, as well as a pharmaceutical composition comprising the TCR, nucleic acid molecule, vector or cell as an active ingredient, and use of the TCR, nucleic acid molecule, vector, cell and pharmaceutical composition in detecting, preventing and / or treating cancers associated with KIT D816Y gene mutation-derived neoantigens.

The present application provides a TCR targeting a FLT3D835H mutation-derived neoantigen, a nucleic acid molecule comprising a nucleotide sequence encoding the TCR or a complementary sequence thereof, a vector containing the nucleic acid molecule, a cell transduced with the nucleic acid molecule or the vector, a pharmaceutical composition comprising the TCR, the nucleic acid molecule, the vector or the cell as an active ingredient, and use of the TCR, the nucleic acid molecule, the vector, the cell or the pharmaceutical composition in detection, prevention and / or treatment of a cancer related to a FLT3D835H gene mutation-derived neoantigen.

The application provides a primer probe set and a kit for detecting a rifampicin-resistant gene of Mycobacterium tuberculosis, and belongs to the technical field of molecular biology detection.The primer probe set comprises RAA primers and qPCR primers targeting the rpoB gene, probe primers targeting the wild-type rpoB gene, and probe primers targeting the mutant rpoB gene.The application adopts the detection methods of RT-RAA and qPCR in sequence, and the above specific primer probe set can be combined to simultaneously and rapidly detect the 516, 526, 531 and 533 sites of the rpoB gene of Mycobacterium tuberculosis, has good specificity, and the detection sensitivity can reach 5 copies / ul, is 20 times higher than the sensitivity of qPCR (100 copies / ul), and has a 5% detection capability for heterogenic drug resistance mutation; the detection method can also greatly shorten the detection time.

This invention provides a crystal form of compound 1, a salt thereof, and a crystal form of the salt thereof. This invention also relates to pharmaceutical compositions comprising the crystal form of compound 1 and the crystal form of a salt of compound 1. This invention further relates to methods of preparing and using these compositions. Compound 1

The invention belongs to the technical field of biology, and provides a method for detecting low-frequency gene mutation based on PNA-PCR combined with a Cas13a system and application of the method. According to the method, a target mutation sequence is enriched through PNA-mediated allele specific amplification, a Cas13a / crRNA system is used for carrying out high-specificity recognition and fluorescence signal amplification on an amplification product, and the high-frequency gene mutation is detected through the Cas13a / crRNA system. High-sensitivity detection of low-frequency gene mutation in plasma circulating tumor DNA (ctDNA) is achieved, and the lower detection limit can reach 0.1%-0.01% VAF. The invention solves the problems of insufficient sensitivity, complex operation, high cost or strong equipment dependence and the like in the prior art such as next-generation sequencing, digital PCR and the like during low-frequency mutation detection, and is suitable for noninvasive rapid screening and precise diagnosis and treatment of clinical tumor gene mutation.

Provided are a gene mutation detection method and apparatus, a device, a medium, and a product. The method includes acquiring a suspected mutation site of a nucleic acid sample under test, where the suspected mutation site is determined based on first mutation feature data generated by a first mutation detection module upon mutation calling performed on sequencing data of the nucleic acid sample under test, and the recall at which the first mutation detection module identifies gene mutation sites is greater than or equal to a preset recall; acquiring second mutation feature data and third mutation feature data of each suspected mutation site; and inputting the second mutation feature data and the third mutation feature data into a pre-trained target mutation detection model and outputting a mutation detection result of each suspected mutation site.

Provided are a fusion protein useful for performing base editing, a method for performing gene editing in a cell at a target site, a dual guide RNA system, and a guide RNA.SOLUTION: To provide fusion proteins and related molecules useful for performing base editing with reduced or no off-target mutations. The fusion protein can comprise a first fragment comprising a nucleobase deaminase or a catalytic domain thereof, a second fragment comprising a nucleobase deaminase inhibitor, and a protease cleavage site between the first fragment and the second fragment. Also provided are improved prime editing systems comprising prime editing guide RNAs with improved stability.SELECTED DRAWING: None

The present invention relates, in part, to chimeric proteins comprising a modified human IFN-γ having one or more mutations that confer reduced affinity and / or biological activity that is restorable by attachment to one or more targeting moieties. The one or more targeting moieties may have recognition domains which specifically bind to antigens or receptors of a tumor or an immune cell. The one or more recognition domains may be a single-domain antibody or a single-chain antibody (scFv). The application also provides use of the chimeric proteins as therapeutic agents.

The application discloses a plant and animal evaluation method based on a genetic algorithm, and the method comprises the following steps: determining initial genes to be evaluated according to evaluation indexes, and determining a chromosome where the initial genes are located as an initial chromosome; setting a preset mutation rate for a position where each gene on the initial chromosome is located, and increasing the preset mutation rate corresponding to the position where the initial gene is located to obtain a target mutation rate; performing genetic simulation on the initial chromosome according to a preset crossover rate, the preset mutation rate and the target mutation rate to obtain a target chromosome; inputting gene coding of the target chromosome into a preset neural network model; determining an evaluation score of the target chromosome in the preset neural network model; and if the evaluation score is higher than a preset threshold, determining that a gene code corresponding to the evaluation score is a high-quality gene. According to the method, genetic simulation is performed in a directional manner according to a target mutation direction, and the problem that the number of iterations is too large and the target solution is not high-quality in the prior art is solved.

Compositions and methods for introducing targeted mutations into the plastid genome are provided.

The invention relates to a genetic variation category prediction method and device based on convex hull geometric constraint, and relates to the fields of bioinformatics, artificial intelligence, applied mathematics and the like, and the method comprises the steps: obtaining multi-modal data corresponding to target genetic variation, including a target variation DNA sequence, a target reference DNA sequence and target semantic text information; performing feature extraction on the multi-modal data through the trained gene variation category prediction model, and predicting the category of target gene variation; wherein loss functions adopted in the model training process comprise a classification loss function used for indicating the difference between a prediction category and a real category, the convex hull geometric constraint loss function is used for indicating the concentration degree of the same type of gene variation corresponding to the distribution range of the feature space and the separation degree of different types of gene variation corresponding to the distribution range of the feature space. According to the invention, based on multi-modal feature fusion and convex hull geometric constraint, efficient and accurate prediction of the gene variation category is realized.

The invention provides a primer and probe combination, a kit and a method for quantitatively detecting the JAK2V617F gene mutation rate, and the primer and probe combination comprises a primer and probe set targeting JAK2V617F mutation sites and a primer and probe set of reference genes GPI, according to the primer probe set targeting JAK2V617F mutation sites, the nucleotide sequence of an upstream primer of a specific mutation site is any one of SEQ ID NO.1-7, the nucleotide sequence of an upstream primer of a specific wild type site is any one of SEQ ID NO.9-14, the nucleotide sequence of a universal downstream primer is SEQ ID NO.8, and the nucleotide sequence of a probe is SEQ ID NO.15. The invention further discloses a kit for detecting the JAK2V617F mutation sites. According to the invention, the JAK2V617F mutation rate can be economically, simply, efficiently, sensitively, accurately and quantitatively detected, and the detection mutation rate is as low as 0.1%.

The application belongs to the technical field of biological detection, and discloses a multiplex signal amplification system for direct detection of DNA / RNA mutation and application thereof, and particularly discloses a multiplex signal amplification system, which comprises an enzyme-labeled plate detection platform, a first signal amplification platform, a second signal amplification platform and a third signal amplification platform; the enzyme-labeled plate detection platform contains a capture nucleic acid probe; the first signal amplification platform contains a nucleic acid probe complementary to a part of the mutation DNA / RNA to be detected; the second signal amplification platform contains two single-stranded DNA probes; the third signal amplification platform contains streptavidin-labeled horseradish peroxidase; and the nucleic acid probe complementary to the part of the mutation DNA / RNA to be detected is a nucleic acid probe containing a hairpin structure. The multiplex signal amplification system can realize detection of a target mutation gene and has good sensitivity and specificity.

A binding protein, comprising a binding domain having antigen specificity for an antigen peptide: HLA complex, wherein the binding domain comprises a T cell receptor (TCR) α chain variable region and a TCR β chain variable region. The use of the binding protein and a pharmaceutical composition comprising the binding protein in the treatment of cancers associated with the antigen peptide.

This application relates to a method, apparatus, computer device, computer-readable storage medium, and computer program product for bacterial phenotypic prediction based on bacterial DNA, which can be used in the field of bioinformatics. The method includes: constructing a heterogeneous graph; the heterogeneous graph includes nodes and the connections between nodes; performing semantic training processing on mutation embedding information based on the heterogeneous graph to obtain mutation semantic embedding information of the mutation embedding information; determining the target mutation semantic embedding information corresponding to the mutation information in the input sample based on the mutation semantic embedding information; performing aggregation processing on the target mutation semantic embedding information to obtain aggregated feature information of the target mutation semantic embedding information; inputting the aggregated feature information into a preset drug resistance prediction model for drug resistance prediction processing to obtain the drug resistance prediction result of the input sample against anti-tuberculosis drugs. This method can improve the efficiency of drug resistance prediction.

A binding protein is provided comprising a binding domain having antigen specificity for an antigen peptide: HLA complex, the binding domain comprising a T cell receptor (TCR) alpha chain variable region and a TCR beta chain variable region. Also provided are uses of the binding protein and a pharmaceutical composition comprising the binding protein in the treatment of cancers associated with the antigen peptide.

This invention relates to a system, kit, and application for the one-pot detection of miRNA and DNA mutations based on CRISPR-Cas12a and aptamers, belonging to the field of biomedical technology. The system for the one-pot detection of miRNA and DNA mutations based on CRISPR-Cas12a and aptamers includes: Cas12a protein, crRNA-R targeting miRNA, aptamer P1, aptamer P2, a linker, AO, and crRNA-D targeting DNA mutations. This invention combines CRISPR / Cas12a with a aptamer-based detection strategy to develop a label-free, direct detection system compatible with smartphones. This system can simultaneously detect EGFR mutations and miRNA-21 expression levels.

The present invention provides a binding protein having antigen specificity for an antigenic peptide / HLA complex, comprising a binding domain containing a T cell receptor (TCR) α chain variable region and a TCR β chain variable region, and uses of the binding protein and pharmaceutical compositions containing the binding protein in the treatment of cancers associated with the antigenic peptide.

The invention belongs to the technical field of gene engineering, and particularly relates to a 3-ketosteroid-delta1-dehydrogenase mutant with improved heat resistance and an application of the 3-ketosteroid-delta1-dehydrogenase mutant. The 3-ketosterone-delta1-dehydrogenase AuKsdD gene derived from arthrobacter urate oxydans is subjected to structural analysis and site-directed mutagenesis design, and a mutant with higher catalytic activity and remarkably enhanced heat resistance compared with a wild type AuKsdD is obtained through directed mutation of key sites E244, K267, T301 and A335. The mutant can keep high catalytic efficiency for a long time under a high-temperature condition, and is suitable for industrial synthesis of steroid drug raw materials.