21 results about "TP53 Genes" patented technology

The invention relates to a method for enhancing target cell removal capacity of immune cells and application. The method comprises the following steps: recombining CD58 and TP53 genes with a granzyme B gene so as to obtain recombinant genes NFAT-GZMB-CD58 and NFAT-GZMB-TP53, and meanwhile, adding a transcription factor NFAT binding site into a regulation region of the obtained recombinant genes. Then, the two recombinant genes are directionally inserted into specific gene loci of immune cells through a gene editing technology. Expression of the two recombinant genes is regulated and controlled by an immune cell activation signal pathway, and translation products of the two recombinant genes can be delivered to target cells of immune cells through a granzyme-perforin pathway, so that the scavenging capacity of the immune cells to the target cells is enhanced, and apoptosis and escape of the target cells are inhibited. The method can be applied to the field of cell therapy.

The invention provides a kit for detecting TP53 gene mutation. The kit for detecting TP53 gene mutation provided by the invention can accurately detect six TP53 gene mutation sites, namely R175H, R249S, R282W, R248Q, R273H and G245S, in genome DNA to be detected at the same time through a multiple fluorescent PCR reaction system and a multi-channel fluorescent signal, can accurately detect the six mutation sites, has ultrahigh amplification efficiency, detection efficiency, sensitivity and specificity, and can be widely applied to detection of TP53 gene mutation. The genotype of a sample with the genome DNA content of 1 ng can be accurately detected, the operation is simple and convenient, and the cost is low.

The invention discloses a mouse gastric adenocarcinoma cell line MPC as well as a construction method and application thereof, and belongs to the technical field of preparation of tumor cell lines and tumor models. The mouse gastric adenocarcinoma cell strain MPC is preserved in China Center for Type Culture Collection, has Cdh1 and Tp53 gene mutations at the same time, has stable cell biological characteristics and typical adenocarcinoma characteristics, can form tumors in immunodeficient and normal immune mice, can be used for preparing a mouse tumor model for occurrence, development and metastasis of gastric cancer, and has a good application prospect. The method is applied to important researches such as molecular biology, cytobiology, detection kit development, new drug research and development and the like.

To provide a new technique related to a multiple myeloma model cell having a deletion in the short arm of chromosome 17.SOLUTION: The method for producing the multiple myeloma-model cells having the deletion in the short arm of the chromosome 17 includes a process for deleting a region containing the whole TP53 genes of the short arm of the chromosome 17 of multiple myeloma-cell line. The multiple myeloma-derived cell strain is one of MM. 1S, H929, MOLP8, and SKMM2, and the step includes a step of introducing a first nucleotide encoding spCas9 into the genome of the multiple myeloma-derived cell strain, and a step of introducing a second nucleotide encoding a plurality of sgRNAs into the genome of the multiple myeloma-derived cell strain, the sgRNAs hybridizing to the short arm of chromosome 17 and having different hybridizing regions.SELECTED DRAWING: None

The objective is to provide a universal method for rapidly detecting cancer-related mutations using dPCR probes. A library is provided for detecting cancer-related mutations in the DNA-binding domain of the TP53 gene, comprising multiple probes and / or primers or primer pairs. This invention enables early diagnosis of post-treatment recurrence of gastrointestinal tumors. Furthermore, it allows for personalized post-treatment follow-up for patients with gastrointestinal tumors.

The present invention concerns novel methods of treatment for venetoclax-resistant acute myeloid leukemia, particularly in subjects with a low expression of TP53 protein or expression of TP53 protein associated with a mutation of the TP53 gene, the methods comprising administering to the subject in need thereof a therapeutically effective amount of a NTRK / ALK / ROS1 inhibitor, or a pharmaceutically acceptable salt thereof.

The invention discloses a liver cancer gene mutation detection method and application, and belongs to the technical field of liver cancer gene detection. The invention finds that deletion mutation of the No.7 exon of the TP53 gene is closely related to the occurrence of liver cancer for the first time, and the research result of the invention can make the early screening sensitivity of liver cancer reach 93.33%, and has important significance for the early detection and intervention of liver cancer. The specific primer pair and the fluorescent probe for digital PCR detection are designed for deletion mutation of the No.7 exon of the TP53 gene, and accurate recognition of specific mutation sites of the TP53 gene is ensured by combining high-fidelity polymerase. According to the invention, the designed detection specific primer is combined with a digital PCR technology, so that the sensitivity of liver cancer gene mutation detection is improved, and the accuracy and reliability of experiments are ensured.

The invention relates to the technical field of gene engineering, and discloses a construction method capable of realizing fluorescence screening of zebrafish tp53 gene knockout homozygote, which comprises the following steps: S1, introducing a knockin plasmid I, Cas9 protein and sgRNA with an antisense strand target sequence as shown in SEQ ID NO: 14 into a zebrafish fertilized egg, and culturing to obtain a strain I; the knock-in plasmid I comprises a left arm, a termination sequence, a right arm and a fluorescence labeling gene I which are arranged in sequence, and the left arm contains a target sequence of sgRNA; s2, replacing the fluorescence labeled gene I in the S1 with a fluorescence labeled gene II, and repeating the S1 to obtain a strain II; and S3, mating the strain I and the strain II, and screening offspring expressing the fluorescent marker genes I and II at the same time. According to the method disclosed by the invention, targeted knockout of the tp53 gene in the zebrafish can be well realized, and the workload of screening and identifying the homozygote can be reduced and the cost can be reduced in the processes of homozygote construction and subculture.

The invention provides a primer pair for detecting TP53 gene exon region mutation and a kit applying the primer pair. The primer pair comprises a first forward primer and a first reverse primer, the nucleotide sequence of the first forward primer is shown as SEQ ID NO: 1, and the nucleotide sequence of the first reverse primer is shown as SEQ ID NO: 2. The primer pair for detecting TP53 gene exon region mutation provided by the invention can be used for performing PCR (Polymerase Chain Reaction) amplification on cDNA (Complementary Deoxyribose Nucleic Acid) obtained by performing reverse transcription on RNA (Ribonucleic Acid) extracted from fresh EDTA (Ethylene Diamine Tetraacetic Acid) anticoagulant whole blood and identifying through Sanger sequencing to find that a PCR amplification product consistent with a theoretical sequence can be obtained; therefore, the specific amplification and mutation detection of the TP53 gene exon coding region with reliable results can be simply, conveniently and quickly completed at one time.

The invention is applicable to the technical field of bioinformatics and molecular diagnosis, and provides a TP53 gene heterozygosity deletion and copy number variation detection method based on targeted sequencing, and synchronous and accurate detection of LOH / CNV and mutation of a TP53 gene is realized through core technology innovations such as customized probe design, dynamic reference set correction, HMM model improvement and KDE double-peak judgment; compared with a traditional detection technology, the problems of CN-LOH leak detection and insufficient integrated analysis of mutation and structural variation are effectively solved, and the coincidence rate with whole exon sequencing reaches 100%; the method has the outstanding advantages of high sensitivity, high specificity, rapidness, high efficiency and controllable cost, can accurately judge the multiple strike states of TP53, provides a reliable molecular diagnosis basis for risk stratification, prognosis evaluation and individualized treatment decision of patients with myeloid tumors such as AML / MDS, and is more suitable for clinical conventional popularization and application.

The application is suitable for the field of bioinformatics and molecular diagnosis technology, and provides a TP53 gene heterozygous deletion and copy number variation detection method based on targeted sequencing. Through core technical innovations such as customized probe design, dynamic reference set correction, improved HMM model and KDE bimodal judgment, the application realizes the synchronous and accurate detection of TP53 gene LOH / CNV and mutation. Compared with traditional detection technology, the application effectively solves the problems of CN-LOH missed detection and insufficient integrated analysis of mutations and structural variations, and the coincidence rate with whole exon sequencing reaches 100%. The method has the outstanding advantages of high sensitivity, high specificity, rapid efficiency and controllable cost, can accurately determine the TP53 multiple hit state, and provides reliable molecular diagnosis basis for risk stratification, prognosis evaluation and individualized treatment decision of AML / MDS and other myeloid tumor patients, and is more suitable for clinical routine popularization and application.

The invention provides a bone marrow smear-based AML standard chemotherapy primary refractory risk prediction method and device. The method comprises the following steps: acquiring a bone marrow smear digital image of an AML patient, processing the bone marrow smear digital image by using an image analysis model to extract morphological characteristics of a cell ecosystem, inputting the morphological characteristics of the cell ecosystem into a TP53 drug resistance estimation model, and estimating the drug resistance of TP53 according to an output result of the TP53 drug resistance estimation model. And predicting whether the primary refractory risk of the TP53 mutant AML of the AML patient is a suspected high risk or not, and outputting prompt information suggested to carry out TP53 gene precise molecular detection under the condition that the primary refractory risk of the TP53 mutant AML of the AML patient is the suspected high risk. Therefore, for a low-risk AML patient, the possibility of TP53 gene precise molecular detection is reduced, the efficiency of predicting the original refractory risk can be improved on the whole, and the problems in the prior art are solved.

The invention provides a primer pair for detecting TP53 gene whole exon region mutation and a kit applying the primer pair. The primer pair comprises a first forward primer and a first reverse primer, the nucleotide sequence of the first forward primer is shown as SEQ ID NO: 1, and the nucleotide sequence of the first reverse primer is shown as SEQ ID NO: 2. The primer pair for detecting TP53 gene whole exon region mutation provided by the invention can be used for carrying out full-length PCR amplification on cDNA (complementary deoxyribonucleic acid) obtained by carrying out reverse transcription on RNA (ribonucleic acid) extracted from fresh EDTA (Ethylene Diamine Tetraacetic Acid) anticoagulant whole blood and identifying through Sanger sequencing to find that a PCR amplification product consistent with a theoretical sequence can be obtained; therefore, the specific amplification and mutation detection of the whole exon region of the TP53 gene can be simply, conveniently and quickly completed at one time, and the result is reliable.

This invention relates to the field of liver cancer biomarker technology. The invention provides a cfDNA gene mutation scoring system to guide liver cancer treatment selection, along with specific biomarkers used: the TP53 gene, AXIN1 gene, VCAN gene, and FBN3 gene. The biomarkers of this invention can classify early-stage liver cancer patients into low-micrometastasis risk subgroups and high-micrometastasis risk subgroups. By performing high-depth capture sequencing on peripheral blood cfDNA isolated preoperatively and detecting mutations in the four genes TP53, AXIN1, VCAN, and FBN3, early-stage liver cancer patients are differentiated into different risk subgroups. Different surgical margins are then selected for different subgroups during liver resection surgery. By using the biomarkers of this invention to classify the risk level of early-stage liver cancer, the selection of surgical margins can be further guided, improving the accuracy of surgical treatment selection for this type of liver cancer and achieving better survival prognosis.

This invention provides a non-invasive artificial intelligence system for predicting EGFR / TP53 co-mutated lung cancer patients, belonging to the field of lung cancer prediction systems. By collecting patients' clinical and imaging characteristics, and using LASSO feature selection and a random forest classifier to build a model, this invention obtains an artificial intelligence prediction system capable of accurately and non-invasively predicting EGFR / TP53 gene co-mutation status in lung cancer patients. Experimental results show that the artificial intelligence prediction system established in this invention has superior predictive performance for EGFR / TP53 gene co-mutation status in lung cancer patients, with an AUC value as high as 0.746 on the test set. This artificial intelligence prediction system provides a new option for clinical screening of lung cancer patients with EGFR+ / TP53+ co-mutations, as well as screening EGFR-mutant lung cancer patients resistant to TKIs, and has important guiding significance for the precise clinical treatment of EGFR / TP53 co-mutated lung cancer patients.

An object is to provide a versatile means utilizing a probe for dPCR, which enables quick detection of a mutation relevant to a cancer. A library including a plurality of probes and / or primers or primer pairs for detecting a mutation relevant to a cancer in the DNA-binding domain of the TP53 gene is provided. By using the present invention, relapse of alimentary canal cancer after treatment can be diagnosed at an early stage. Individualized post-treatment follow-up of an alimentary canal cancer patient is also enabled.

The application provides a kit for detecting TP53 gene mutation. The kit for detecting TP53 gene mutation can accurately detect six TP53 gene mutation sites (R175H, R249S, R282W, R248Q, R273H and G245S) in the to-be-detected genomic DNA through a multiplex fluorescence PCR reaction system and a multi-channel fluorescence signal, and can accurately detect the six mutation sites, has ultrahigh amplification efficiency, detection efficiency, sensitivity and specificity, can accurately detect the genotype of a 1 ng sample of genomic DNA, and is simple to operate and low in cost.

The invention relates to the field of cancer, in particular breast cancer. In particular it relates to the field of immune system directed approaches for tumor reduction and control. Some aspects of the invention relate to vaccines, vaccinations and other means of stimulating an antigen specific immune response against a tumor in individuals. Such vaccines comprise neoantigens resulting from frameshift mutations that bring out-of-frame sequences of the GATA3, CDH1, MAP3K1, RUNX1, and TP53 genes in-frame. Such vaccines are also useful for ‘off the shelf’ use.

The invention relates to a detection method and a kit for detecting TP53 gene mutation based on dPCR, primers and probes of the detection method are designed according to a TP53 gene DNA sequence, a pair of primers and two Taqman probes are respectively designed aiming at 6th and 8th exon gene sequences of the TP53 gene, the sequences of the primers and the probes are the same as the sequence of a wild type TP53 gene, and the TP53 gene mutation can be detected by the detection method and the kit. The two probes on each exon are mutually referenced, and when mutation such as insertion, replacement and deletion exists in a probe coverage area, the corresponding probe cannot be combined with a template and is subjected to enzyme digestion to emit fluorescence, so that the probe can detect various mutation of the TP53 gene. The detection method comprises the following steps: extracting sample DNA, preparing a dPCR reaction mixed solution, preparing micro-droplets, carrying out PCR amplification, collecting signals and calculating the concentration. The kit for detecting TP53 gene mutation is easy to operate, high in sensitivity, low in price and suitable for continuous monitoring of mutation cloning loads of patients.

The application provides a construction method and application of a mouse spontaneous tumor model of gastric cancer, and the construction method comprises the following steps: reducing or inhibiting the expression amount of a TP53 gene and / or the activity of a P53 protein in a mouse, and then chemically inducing the mouse by using a drug to obtain a mouse spontaneous tumor model of gastric cancer; wherein the drug comprises sodium nitrite and ethyl sarcosine, and / or the drug comprises an N-nitroso compound. The application first proposes to use NESS to construct a spontaneous gastric cancer model of a C57 mouse, and the construction method of the mouse spontaneous tumor model of gastric cancer provided by the application has a tumor formation rate of 75% or more; the tumor formation cycle is short, about 3-4 months, and the construction cost is low.

The present invention relates to the use of POLR2A as a biomarker for the efficacy of treating tumors. The inhibitory effects of histone deacetylase inhibitors and echinocandin antifungal drugs on tumor cells with low POLR2A expression levels are significantly higher than those on tumor cells with high POLR2A expression levels. In tumor cells with low POLR2A expression levels, there is hemizygous deletion of the POLR2A gene; and / or the expression product of the POLR2A gene has a lower expression level compared to the control; and / or there is hemizygous deletion of the TP53 gene. This compound can effectively predict the efficacy of histone deacetylase inhibitors in treating breast cancer, colon cancer, prostate cancer, and acute leukemia, providing a new method for the precise treatment of cancer and thus avoiding the blindness of chemotherapy.