339 results about "Enhancer" patented technology

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The invention provides a kit and a method for detecting common fusion genes of lymphoid leukemia based on a multiple digital PCR (Polymerase Chain Reaction) method. By constructing a synergistic effect mechanism of the primer and the fluorescent probe and optimizing matching of the primer, the probe and a fluorescent marker, non-specific binding among amplification products is effectively avoided. A composite reaction aid with a specific formula, namely a PCR enhancer, is innovatively introduced, so that the amplification efficiency and specificity of a multiple detection system are remarkably improved. According to the kit provided by the invention, the integrated detection capability on seven fluorescent channels, namely Atto 425, VIC, FAM, ROX, CY5, CY5.5 and CY7, is realized in a single-hole design. By innovatively constructing an ROX + CY5 and Atto425 + VIC dual-channel joint detection system, when the ROX and CY5 channels or Atto425 and VIC channels in the same detection hole simultaneously present positive signals, two additional detection targets can be specifically interpreted. The innovative detection strategy based on channel combination enables the kit to realize precise detection of 18 fusion genes only through two detection holes in a breakthrough manner.

The present disclosure relates to compositions and methods for reducing expression of MYC gene in a cell. In some embodiments, an expression repressor comprises a targeting moiety that binds a MYC promoter, anchor sequence, or super-enhancer. In some embodiments, the expression repressor comprises an effector moiety that represses transcription or methylates DNA. Systems comprising two expression repressors are also disclosed. The compositions can be used, for example, to treat cancers such as HCC.

The invention relates to the technical field of cancer risk prediction, in particular to a deep learning cancer risk prediction method and system based on a methylation sequence. The method comprises the following steps: acquiring DNA methylation sequencing data and reference genome annotation data in a blood sample of a cancer patient; performing functional region classification processing on the DNA methylation sequencing data according to the reference genome annotation data to obtain promoter subregion methylation data, enhancement subregion methylation data and transcription factor binding site methylation data; performing dynamic sequence similarity calculation processing on the methylation data of the promoter region to obtain a first feature vector containing importance weight of the promoter region of the cancer-related gene; the methylation sites are classified according to the functions of the methylation sites in the transcriptional regulation pathway, so that the biological interpretation of the characteristics is improved, and the synergistic effect among different functional regions can be captured.

The invention provides an ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting an anti-MDA5 antibody and a preparation method of the ELISA kit. According to the kit, full-length human MDA5 protein (1025 amino acids) expressed by a cell-free protein expression system under the low-temperature condition of 22 DEG C is adopted as a coating antigen, and a disulfide bond enhancer E6820 is added into the system to promote correct folding of the protein. The kit comprises a microwell plate coated with a full-length MDA5 antigen, an enzyme-labeled anti-human IgG secondary antibody, a standard substance with the concentration range of 1-243 ng / mL, a chromogenic substrate and an optimized confining liquid system. The invention solves the problem of insufficient sensitivity caused by the use of fragment antigens in the prior art, and can be used for early diagnosis and illness monitoring of dermatomyositis-related interstitial lung diseases.

The present invention relates to an exogenous target gene expression cassette for delivering an exogenous target gene to the retina, in particular AIPL1 to retinal pigment epithelial cells and photoreceptor cells, comprising an IRBP enhancer sequence, a rhodopsin kinase (RK) promoter sequence and a CAG intron sequence, which are operatively linked, and an exogenous target gene. The present invention also relates to a recombinant adeno-associated viral vector comprising a viral capsid comprising a capsid protein or a capsid protein variant and a viral vector genome comprising an expression cassette encoding for specifically expressing an exogenous target gene in retinal pigment epithelial cells and photoreceptor cells. The recombinant adeno-associated virus vector can be used for relieving or treating retinal degenerative eye diseases by intravitreal administration or subretinal administration.

The present disclosure provides engineered immune cells modified to overexpress tumor sensing enhancer proteins, thereby providing the engineered immune cells with an enhanced ability to migrate to and / or infiltrate tumors. In some examples, the tumor sensing enhancer proteins are tumor sensing receptors that specifically bind to non-chemokine receptors. The disclosure also provides methods including the provided engineered immune cells.

Use of PGLYRP2 gene and a related drug thereof. The present invention relates to the field of biopharmaceuticals, and in particular, to use of the PGLYRP2 gene and the related drug thereof. The present invention aims to solve the problem that existing drugs can only inhibit the replication of hepatitis B virus and cannot eliminate viral cccDNA. The PGLYRP2 gene is used for preparing a drug for treating a hepatitis B virus infection. The PGLYRP2 gene is used for preparing a drug for eliminating hepatitis B virus cccDNA. Also provided is a gene therapy vector for eliminating hepatitis B virus. The gene therapy vector comprises a promoter, an intron, an enhancer, and a PGLYRP2 gene encoding region. The HBV virus inhibition function and the cccDNA-eliminating effect of the PGLYRP2 protein are defined, so that the elimination of HBV in liver cells and a mouse liver can be significantly promoted.

The invention discloses an ensemble learning and deep learning-based enhancer prediction method, which comprises the following steps of: firstly, performing enhancer prediction on multi-dimensional epigenetic signal data by utilizing a Blending-KAN model so as to identify an enhancer region, and on the basis, aiming at the region which is predicted as the enhancer, segmenting the region into subsequences through a sliding window so as to obtain an enhanced region; the method comprises the following steps: firstly, constructing a plurality of sub-sequences, further predicting the probability that each sub-sequence is an enhancer through a Stacking-Auto model, and finally, accurately positioning a complete enhanced sub-region by adopting a dynamic threshold algorithm based on the probability values; according to the multi-model combination method provided by the invention, the accuracy and fine positioning capability of enhancer identification are effectively improved, and the method is expected to be applied to more complex gene regulation network researches.

The present invention relates to engineered liver-specific core promoters, synthetic promoters (which contains the engineered core promoters and enhancers), expression vectors (which contains the synthetic promoter), as well as methods of using the promoter or the expression vector thereof to address the need in the field, including treatment of various genetic diseases or conditions associated with the liver. In some embodiments, the liver-specific promoter includes continuous or discontinuous genome sequences from SERPINA1 genome.

The invention provides a kit for detecting human BRAF gene V600E and TERT gene promoter mutation, and belongs to the technical field of biology. The kit comprises a primer, a probe, a 5 * PCR (Polymerase Chain Reaction)-Enhancer, a 10 * PCR Buffer, dNTP (Deoxyribonucleoside Triphosphate) Mix, MalTaq DNAPlyase, magnesium ions, DEPC (Diethylpyrocarbonate) water, a positive control and a negative control. According to the present invention, the ARMS-PCR detection technology is adopted to detect the specific gene mutation, and the target mutation site is identified by designing the specific primer and the probe, such that the detection specificity and the detection sensitivity are improved; by combining with a Taqman probe technology, the fluorescent signal change in the PCR amplification process can be monitored in real time, so that whether mutation of a BRAF V600E site, a TERT C228T site and a TERT C250T site exists or not is accurately judged; according to the method, pre-amplification of 15 cycles is adopted, then quantitative amplification of 30 cycles is adopted, and when the initial amount of the target DNA is very low, the copy number of the target DNA can be increased, so that the detection accuracy and sensitivity are further improved.

The invention relates to the field of biological medicine, and particularly discloses a regulatory element combination for driving nucleic acid expression based on ITR-enhancer and application of the regulatory element combination. The regulatory element combination for driving nucleic acid expression based on an ITR-enhancer comprises at least one inverted terminal repeat (ITR) and at least one enhancer, the regulatory element combination is delivered through an AAV vector. The invention also provides a recombinant nucleic acid molecule for improving specific delivery of the target gene. The recombinant nucleic acid molecule comprises the regulatory element combination, an exogenous target gene and polyadenylic acid. The AAV vector provided by the invention can be used for driving the expression of a target gene without the action of a traditional promoter, and has the advantages of enhancer tissue specificity, short sequence (50-100bp), pathological microenvironment response and the like, thereby breaking through the capacity, targeting and dynamic limitation of the traditional AAV vector, improving the DNA (Deoxyribose Nucleic Acid) carrying capacity of the AAV vector, and simultaneously improving the expression of the target gene. The tissue and spatio-temporal expression specificity of a target gene is realized.

The present invention relates to chimeric promoters without CpG motifs. The chimeric promoter comprises a CMV enhancer, an IFN-gamma gene core promoter and an intron sequence, and a CG motif in the CMV enhancer, the IFN-gamma gene core promoter and the intron sequence is subjected to substitution mutation. The promoter can improve the expression efficiency of exogenous genes in activated immune cells.

To provide a method for simply and efficiently evaluating the inducibility of a foreign matter response reaction in the skin of a test sample without using an animal, and to provide a substance having the inhibitory properties of the foreign matter response reaction in the skin and a method for searching the same.SOLUTION: The method includes a step of bringing keratinocytes transformed with a vector having an enhancer sequence containing a foreign body-responsive sequence and a sequence encoding a reporter protein downstream of the enhancer sequence into contact with a solution containing a test sample, a step of measuring the expression level of the reporter protein in the keratinocytes, and a step of evaluating the inducibility of a foreign body response reaction of the test sample in the skin based on the expression level of the reporter protein. The problem is solved by providing an agent for suppressing foreign matter response reaction in the skin containing fregula 1 found by the search method.SELECTED DRAWING: Figure 3

The invention provides an artificial plant bidirectional constitutive strong promoter, and particularly relates to a high-activity short transcription enhancer fragment which is formed by connecting three plant endogenous fragments and a plant virus in series and serves as a transcription activation basic element. A minimum core promoter element Mini 35S from a cauliflower mosaic virus and a minimum core promoter element CsVMV mini-pro functional module from a cassava vein mosaic virus are respectively integrated at two ends, so that the plant constitutive strong promoter with bidirectional starting capability is constructed. According to the invention, a synthetic quadruple STE transcription enhancer, a Mini 35S promoter and a CsVMV mini-pro promoter are functionally coupled to form an expression regulation tool with broad-spectrum adaptability to plant species, and two operably connected target gene expression cassettes can be simultaneously driven by a short regulation sequence to realize synergistic efficient expression in plant cells.

The invention discloses an enhancer regulatory element for transcriptional regulation expression of a human LTF gene and application of the enhancer regulatory element. The sequence of the regulatory element is SEQ ID No.1. Experiments prove that the human specific regulatory element can significantly activate report gene expression (improved by 14.93 times) in a bovine mammary epithelial cell line (MACT), the limitation of dependence on species conservative elements in the prior art is broken through, and an expression system designed based on the element can realize human level LTF high-efficiency expression. And a brand new tool is provided for development of mammary gland bioreactors and functional dairy products.

The invention belongs to the technical field of gene engineering, and particularly relates to a medicine for treating AML (acute myelogenous leukemia) and / or prognosis of AML patients and application of ubiquitin specific protease 20 serving as a target spot in treating acute myelogenous leukemia. The invention provides a medicine for treating AML (acute myelogenous leukemia) and / or prognosis of an AML patient and application of ubiquitin specific protease 20 as a target spot in treating acute myelogenous leukemia, USP20 is used as a super enhancer regulation gene, and the progress of the AML is promoted by combining with CTNNB1, ERG, ELF1 and RUNX1. The knock-down of the USP20 can significantly inhibit AML proliferation in vivo and in vitro. The wnt-beta-catenin pathway can be influenced by interfering the expression of the USP20 so as to influence the progress of AML (acute myeloid leukemia). And the inhibition effect of the inhibitor AS1517499 subjected to virtual screening on the growth of the AML cells is superior to that of a commercial inhibitor GSK2643943A of USP20.

Described herein is an oncolytic herpes simplex virus, G47ΔhIL12A, which is G47Δ containing a cassette expressing a transgene, e.g., human IL-12, driven by a spontaneously arising genetically altered HCMV immediate-early (IE) enhancer / promoter. This virus has augmented (A) production of the transgene and increased virus replication while retaining safety. Also provided are methods of use thereof for treating cancer, e.g., glioblastoma (GBM) and triple-negative breast cancer (TNBC).

The present invention relates to an adeno-associated virus (AAV) vector comprising in the 5' to 3' direction: a first AAV2 ITR sequence; a CMV enhancer sequence; a chicken β-actin promoter sequence; a nucleic acid sequence encoding an apolipoprotein 2 (APOE2) polypeptide comprising a Christchurch mutation; a polyA sequence; and a second AAV2 ITR sequence.

The present invention provides methods and compositions for treating cancer in a subject comprising the use of therapeutic cells and compositions to prevent or reduce T cell exhaustion. Modified hematopoietic cells comprising a genomic modification in an enhancer surround the GAB3 gene are also provided.

In one aspect, a method of treating an NK cell to induce a gene edit. The method may include contacting an NK cell with a transfection composition including poly-L-glutamic acid (y- PGA), a single-stranded oligo deoxynucleotide (ssODN) enhancer, or a combination thereof; contacting the NK cell with an editing composition including: a DNA-PK inhibitor, and an HD AC inhibitor; electroporating the NK cell; and contacting the NK cell with a recovery composition including DNase I. In another aspect, a method of preparing an engineered NK cell comprising a fluorescent reporter at an endogenous locus. In another aspect, a method of treating an NK cell to introduce an exogenous nucleic acid.

Disclosed herein is a chimeric nucleic acid including a human telomerase reverse transcriptase (hTERT) enhancer or a fragment thereof and an SV40 enhancer or a fragment thereof and methods of using the same.

The invention belongs to the technical field of biomedical detection, and discloses a TAR DNA binding protein 43 detection method based on LC-MS / MS and application. The detection method comprises the following steps: taking a biological matrix to be detected, and filtering to obtain filtrate; adding an ion enhancer into the obtained filtrate, oscillating and incubating, adding a protein precipitant, and separating precipitated protein; adding trypsin into the precipitated protein, and incubating to obtain a sample containing a characteristic peptide fragment; and detecting the TAR DNA binding protein 43 in the sample by adopting liquid chromatography-triple quadrupole mass spectrometry. According to the method, the TDP-43 is specifically detected and quantified by adopting a liquid chromatography-tandem mass spectrometry technology, so that the excellent effects of efficiently detecting and accurately quantifying the TDP-43 and improving the detection sensitivity and stability are achieved. The method can be applied to the fields of in-vitro diagnosis of neurodegenerative diseases, pathological mechanism research and therapeutic drug development.

A duchenne muscular dystrophy-related exonic splicing enhancer, sgRNA and gene editing tool can be applied in the preparation of drugs for treating duchenne muscular dystrophy. The gene editing tool designed on the basis of cytosine deaminase AID mutants and Cas9 mutants can perform site-specific modification on a mammalian genome by using an adeno-associated virus (AAV) as a vector. By optimizing an encoding nucleic acid sequence and an element composition structure of the editing tool, site-specific targeted modification of mammalian genetic material DNA can be efficiently achieved; and by performing targeted genetic manipulation on the nucleic acid sequence carrying disease mutations, a pathogenic mutation cannot be retained in a mature protein amino acid sequence or the pathogenic mutation cannot perform its function, so that the purpose of treating various gene mutation type genetic rare diseases is achieved, and the advantages of high efficiency, safety and stability are achieved.

The invention belongs to the technical field of gene editing, and particularly relates to a WER10 enhancer and application thereof. The invention provides application of a WER10 nucleic acid molecule and / or a biological material for regulating and controlling the WER10 nucleic acid molecule, the nucleotide sequence of the WER10 nucleic acid molecule is as shown in SEQ ID NO: 1, and the application comprises any one or more of the following items: 1) application of the sequence as shown in SEQ ID NO: 1 as an enhancer; 2) application in enhancing the expression of a downstream reporter gene LUC induced by an immune induction signal; and 3) application in regulation and control of the broad-spectrum antibacterial property of the plants. The sequence and the characteristics of the pathogenic bacterium induced enhancer WER10 located in the intergene region are identified and verified from arabidopsis thaliana, plant immunity related cis-regulatory element resources are enriched, and a new resource is provided for constructing a pathogen induced disease-resistant expression system and developing crop disease-resistant molecular design and breeding.

The invention relates to the technical field of biology. Specifically, the invention relates to an enhancer and a promoter for eye gene therapy, and a recombinant adeno-associated virus vector containing the enhancer and the promoter. The invention also relates to a pharmaceutical composition comprising the recombinant adeno-associated virus vector and its use in the treatment of glaucoma.

The present invention discloses a method for screening non-small cell lung cancer markers based on enhancer methylation differences, as well as the markers and applications thereof. Abnormal DNA methylation in the enhancer regions of the human genome can lead to abnormal gene expression regulation in non-small cell lung cancer. The method for screening non-small cell lung cancer markers described in the present invention is based on this principle and further combines paired-end tag sequencing analysis of chromatin interaction technology (ChIA-PET) to screen out DNA methylation markers in the genomic enhancer regions that cause abnormal gene expression regulation in non-small cell lung cancer. This screening method has the advantages of accuracy, rapidity, and high throughput. The DNA methylation markers described in the present invention are cg00787780, cg16434331, cg21862081, and cg24327132. The markers have good diagnostic efficacy for non-small cell lung cancer, showing high accuracy, high sensitivity, and strong specificity, and can better help improve the detection rate of non-small cell lung cancer.

Genome editing systems, guide RNAs, and CRISPR-mediated methods are provided for altering portions of the HBG1 and HBG2 loci, portions of the erythroid specific enhancer of the BCL11A gene, or a combination thereof, in cells and increasing expression of fetal hemoglobin.

Provided is a promoter having high activity in an activated T-cell. The promoter comprises, from 5′-end to 3′-end, a CMV enhancer, an IFNγ promoter, and a long terminal repeat sequence from human T-cell leukemia virus that are connected in sequence. The promoter exhibits greater activity in an activated immune cell than the existing promoters and is low in activity or inactive in other non-immune cells.

The invention relates to the technical field of aquatic animal immune regulation and control, in particular to rHoLL-like stepped lectin-like recombinant protein as well as a preparation method and application thereof. The recombinant protein has pathogen-related molecular pattern recognition and combination capabilities, can promote aggregation of pathogens in a body fluid environment, and activates an aquatic animal complement system, so that the non-specific immune defense capability is enhanced. The preparation method comprises the following steps: amplifying a HoLL-like mature peptide coding sequence, constructing a recombinant expression vector, and converting the recombinant expression vector into an escherichia coli expression host for induced expression to obtain rHoLL-like lectin-like recombinant protein existing in an inclusion body form; and carrying out affinity chromatography purification on the inclusion body protein, and carrying out renaturation treatment in a manner of gradient reduction of denaturant concentration to obtain the rHoLL-like stepped lectin-like recombinant protein. The rHoLL-like stepped lectin-like recombinant protein has good biological safety, and can be applied to aquaculture as an aquatic animal immunopotentiator or a related biological product.