24 results about "Luciferase" patented technology

This invention relates to the packaging and screening of viral vectors, particularly to the packaging and screening of AAV mutants, specifically an adeno-associated virus mutant that efficiently infects HT-22 cells. By constructing a peptide mutant library of AAV9, a new AAV9 mutant with seven inserted amino acids was obtained through screening and verification. This mutant can effectively infect HT-22 cells at an MOI of 1E+5, achieving a higher infection rate than the natural AAV9 serotype at MOI=1E+5. The AAV9-HT01 mutant obtained after screening showed the best effect, with a significantly higher infection positivity rate compared to the control AAV9 serotype under the same MOI conditions. The fold increase was 4.4-fold as measured by luciferase (RLU) value detection. This effectively reduces the amount of AAV cells used for infection, saving experimental costs, and simultaneously makes it feasible to study the mechanisms of gene therapy for central nervous system diseases in HT-22 cells.

The application belongs to the technical field of biotechnology, and particularly relates to a colorectal cancer high liver metastasis cell line and a preparation method and application thereof. The cell line was preserved in the China Center for Type Culture Collection on January 14, 2026, and the preservation number is CCTCC NO: C202618. The cell line is derived from a mouse colorectal cancer cell line MC38, and is constructed by lentivirus transfection to express luciferase stably, and is obtained by continuously performing at least five rounds of liver metastasis tumor orthotopic iteration screening in C57BL / 6 mice through rectal submucosal injection. The MC38-P06 cell line provided by the application has a significantly enhanced liver metastasis ability, a shorter MC38-P01 model time, a higher liver tumor load, and a shorter mouse survival period, and can be used for screening and evaluating anti-liver metastasis drugs, researching liver microenvironment regulation mechanisms, and identifying liver metastasis related diagnostic markers.

The present application relates to in situ model modeling method, especially for a kind of non-invasive cervical cancer in situ model modeling method of mouse, including selecting several 8 weeks old BALB / c-nude mouse, all mice are divided into non-invasive group and operation group, the age, weight of two groups of mice are compared, and there is no statistically significant difference;Non-invasive group is widened with the cotton stick of diameter 3mm and wipes dry vagina, the tumor block with luciferase fluorescence is immersed in modified hydrogel, after heating and making modified hydrogel solidification, the tissue glue mixed with hard gypsum in modified hydrogel is dropped on the surface of modified hydrogel and quickly inserted into vagina, when there is obstruction, stop deepening, pause 30s and wait for tissue glue to be completely dry, when tumor block is fixed in cervix, it is modeling completion;Operation group first opens abdominal cavity, and tumor tissue of same size is sutured on cervix.This kind of non-invasive cervical cancer in situ model modeling method of mouse, using non-invasive method to replace operation method to establish non-invasive cervical cancer in situ model, can reduce the pain of mouse, reduce experimental difficulty.

The present invention relates to the packaging and screening of viral vectors, and in particular to the packaging and screening of AAV mutants, specifically an adeno-associated virus mutant suitable for specific infection of rat hepatocytes. By constructing an AAV9 peptide mutation library, a new AAV9 mutant with 7 amino acids inserted is obtained through screening and verification. The mutant can effectively infect BRL cells under an MOI of 1E+5, achieving an infection effect higher than that of the natural AAV9 serotype MOI of 1E+5. The AAV9-BRL04 obtained through screening has the best effect. Under the same MOI conditions, the infection positive rate is significantly improved compared with the control AAV9 serotype. The multiple is statistically 8 times as high as detected by luciferase (RLU) value, which effectively reduces the amount of AAV-infected cells used, saves experimental costs, and makes it feasible to conduct gene therapy-related mechanism research in rat hepatocytes.

The invention belongs to the technical field of gene engineering, and relates to a construction method and verification of a fixed-point knock-in mouse model expressed by fluorescent tracing IFN-gamma protein. According to the invention, a luciferase fluorescent gene is inserted in front of a 3 'UTR (Untranslated Region) of an IFN-gamma gene in a homologous recombination manner by utilizing a CRISPR / Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR associated 9) technology, and connection is carried out through 2A oligopeptide, so that an IFN-gamma-2A-luciferase (IFNG-Luc for short) fluorescent tracing transgenic mouse model is constructed; the model can be used for realizing the co-expression of IFN-gamma (interferon-gamma) and luciferase protein. By utilizing the model, the expression position of the IFN-gamma protein can be marked through the luciferase fluorescence expression, and the expression intensity of the IFN-gamma protein can be reflected through the luciferase fluorescence intensity, so that the effect of the IFN-gamma in specific physiological and pathological processes can be researched.

This invention discloses the application of the apple C2H2 zinc finger protein transcription factor MdZAT1 gene in anthocyanin synthesis, belonging to the field of plant genetic engineering technology. This invention isolates a C2H2 zinc finger protein transcription factor MdZAT1 gene from apples and experimentally reveals a negative correlation between MdZAT1 gene expression and anthocyanin content in apple fruit during development. Overexpression of MdZAT1 in apple callus tissue inhibits anthocyanin synthesis; transient overexpression and silencing experiments in apple peel show that MdZAT1 is a negative regulator of anthocyanin accumulation in apple peel. Yeast one-hybrid assays, EMSA, and dual-luciferase assays verify that MdZAT1 protein binds to and negatively regulates MdMYB114, MdCHI, and MdANS. The MdZAT1 gene has practical application value in regulating anthocyanins in apple fruit, providing a basis for improving the appearance and quality of apple fruit.

This invention provides a nitro-aryl ring-substituted imidazopyrazine compound that can be used as a bioluminescent probe for detecting nitroreductases. In the NanoLuc luciferase bioluminescent system, this probe exhibits good responsiveness in in vitro experiments and can be selectively reduced by NTRs. Viable bacterial experiments and imaging experiments in a mouse model of bacterial infection both demonstrate that this probe is a highly selective and sensitive luminescent probe for detecting NTR activity, enabling the detection and imaging of NTR activity in vivo.

This invention relates to the pharmaceutical field, specifically to the application of tissue inhibitor metalloproteinases-2 (TIMP2) in the preparation of drugs for the prevention or treatment of traumatic brain injury. TIMP2 protein can increase the fall latency in rotarod experiments in mice with traumatic brain injury; improve the motor balance ability of mice with traumatic brain injury on a balance beam; improve neurological function impairment in mice with traumatic brain injury; and reduce Evans blue permeability in brain tissue. In brain microvascular endothelial cells (HBMECs), TIMP2 protein can participate in maintaining the integrity of the endothelial barrier, reversing the loss of expression and altered localization of the connectome complex induced in an in vitro traumatic brain injury model, and reducing luciferase leakage. This invention also provides the application of TIMP2 protein as an Integrin α3β1 ligand in the preparation of drugs for the treatment of central nervous system diseases caused by blood-brain barrier dysfunction. TIMP2 protein exerts its function by binding to the cell membrane receptor Integrin α3β1, regulating VE-Cadherin phosphorylation. TIMP2 protein shows promising application prospects in the treatment of blood-brain barrier dysfunction caused by traumatic brain injury.

The invention relates to the technical field of medicinal chemistry, in particular to application of hovenia acerba total triterpenes to preparation of a medicine with a non-small cell lung cancer resisting effect. Researches show that the hovenia acerba total triterpenes have the effects of resisting NSCLC and enhancing sensitivity; the hovenia acerba total triterpenes can inhibit A549 cell proliferation, migration and apoptosis in a dose-dependent manner, and can stimulate the formation of autophagosome and induce an autophagy process, which indicates that the hovenia acerba total triterpenes have the effect of KNSCLC. The hovenia acerba total triterpenes can inhibit the activity of ARE-Leu luciferase and increase the cytotoxicity of the hovenia acerba total triterpenes combined with cisplatin, and the hovenia acerba total triterpenes combined with cisplatin can significantly reduce the protein expression of the Nrf2 pathway, which shows that the hovenia acerba total triterpenes have the effect of enhancing the sensitivity of NSCLC drug-resistant cells. Therefore, the hovenia acerba total triterpenes used as the active ingredient for preparing the medicine with the anti-NSCLC effect have important application value.

The present application relates to a kind of genetic code NAD+ protein probe based on resonance energy transfer and its preparation method and application, specifically disclose a kind of genetic code NAD+ protein probe, it is formed by resonance energy transfer donor, NAD+ response protein and resonance energy transfer acceptor series connection;Wherein NAD+ response protein is the mutant of DNA ligase, the sequence of the mutant of DNA ligase is as shown in SEQ ID NO.3, or SEQ ID NO.6;The resonance energy transfer donor is selected from luciferase or fluorescent protein;The resonance energy transfer acceptor is selected from fluorescent protein and the fluorescent protein as resonance energy transfer acceptor is different from the fluorescent protein of resonance energy transfer donor.The protein probe of the present application can be synthesized in living cell and be used to detect NAD+ concentration in living cell.

The invention discloses an amylose improvement method based on Wx gene intron branch point editing and application of the amylose improvement method, and relates to the technical field of plant genetic engineering. The method comprises the following steps: (1) carrying out biosignal analysis to obtain a peripheral sequence of a rice intron branch point (BS); (2) preliminarily determining the influence generated by target deletion through a dual-luciferase experiment; (3) carrying out agrobacterium transformation to obtain a mutant strain; (4) determining that Wx expression of the intron mutant has variation by utilizing qPCR (quantitative polymerase chain reaction) and SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis); and (5) rice quality test results show that the knockout of the 5 # intron and the 7 # intron can change the function of the Wx gene, so that the amylose content of the rice is reduced, and the gel consistency is increased. The method can be applied to cultivation of new varieties and / or new strains of rice with low amylose content and / or high gel consistency, and is of great significance to production of high-quality rice.

The invention discloses a hippocampus extract with activity of improving renal anemia as well as a preparation method and application of the hippocampus extract, and belongs to the technical field of biological medicines. The hippocampus extract with different molecular magnitudes is prepared through optimization of a membrane separation process, experiments prove that the hippocampus extract can remarkably improve the activity of fluorescein enzyme and promote generation of EPO of a human embryonic kidney cell HEK293 cell line, and particularly, the blood enriching activity of the hippocampus extract with the molecular weight smaller than 20 kDa is obviously higher than that of a hippocampus total extract, so that the hippocampus extract with the molecular weight smaller than 20 kDa has a good application prospect. The traditional Chinese medicine composition can effectively reduce the inosine and urea nitrogen levels of rats suffering from renal injury type anemia and improve renal failure, and has the effect of remarkably improving renal anemia. The hippocampus extract obtained through optimization of the preparation method relieves the symptoms of renal anemia from multiple aspects, improves renal injury, provides a scientific basis for clinical research of drugs for improving renal anemia and development of functional foods, and has great application value.

The invention provides a cDNA (complementary deoxyribonucleic acid) clone plasmid of an avian infectious bronchitis virus (IBV) for expressing secretory luciferase as well as a virus strain and a vaccine obtained by rescuing, and belongs to the technical field of biological medicines. The invention provides a construction method of cDNA (complementary deoxyribonucleic acid) clone plasmids of IBV (infectious bursal virus), which is completed by a one-step method based on yeast homologous recombination in a segmented amplification mode. The IBV infectious cDNA clone constructed by the method has good stability, an IBV genome can be transformed on a gene level by directly utilizing in-vitro homologous recombination, and the efficiency of constructing a recombinant virus is improved. According to the invention, a non-essential gene of the infectious cDNA clone is replaced by a Gluc reporter gene, and the obtained infectious cDNA clone directly transfects mammalian cells to rescue recombinant viruses. The recombinant virus is constructed through a virus rescue method, the growth characteristic of the recombinant virus is basically consistent with that of a parent virus, and genetic stability is kept.

The invention relates to the field of plant genetic engineering, in particular to application of an MYB transcription factor gene HvPHL2 in regulating and controlling the content of beta-glucan in barley grains. The MYB transcription factor gene HvPHL2 participating in regulation and control of the barley grain beta-glucan content is found, and through construction experiments of yeast single impurity, gel migration, dual luciferase and overexpression genetic materials, it is proved that the transcription factor HvPHL2 can be combined to CCAAT-box of a promoter region of a barley grain beta-glucan synthesis major gene HvCslf6, expression of an HvCslf6 promoter is inhibited, and the content of the barley grain beta-glucan in the barley grain beta-glucan synthesis major gene HvCslf6 in the barley grain beta-glucan synthesis major gene HvCslf6 in the barley grain beta-glucan synthesis major gene HvCslf6 is regulated and controlled. The method plays a key role in regulation and control of beta-glucan synthesis, and provides a new theoretical basis for genetic regulation and control of the beta-glucan content of the barley grains.

The present application relates to the packaging and screening of viral vectors, in particular to the packaging and screening of AAV mutants, specifically an adenovirus associated virus mutant specifically infecting L6 cells; the present application constructs a peptide segment mutation library of AAV9, and obtains a new AAV9 mutant with 7 inserted amino acids through screening verification, the mutant can effectively infect L6 cells under the condition of MOI=1E+5, and the infection effect is higher than that of the natural AAV9 serotype under the condition of MOI=1E+5, the effect of AAV9-L601 obtained through screening is the best, under the same MOI condition, the infection positive rate is obviously improved compared with the control AAV9 serotype, the multiple of the luciferase (RLU) value is 10.8 times through detection and statistics, the use amount of AAV infected cells is effectively reduced, and the experimental cost is saved.

The invention belongs to the technical field of biomedicine, and particularly relates to application of hydroxytyrosol in liver organ function optimization. The invention discloses a vomitoxin antagonist screening method based on a TGR5 target spot. The vomitoxin antagonist screening method comprises the following steps: constructing a TGR5 / pCREB tool cell; screening a natural product by combining a luciferase luminescence kit; processing the tool cells; if the luminous intensity is obviously increased, the natural product has the function of activating the TGR5, and if the luminous intensity is reduced, the natural product has the activity of inhibiting the TGR5. According to the application disclosed by the invention, the hydroxytyrosol is found to simultaneously improve liver cell functions and liver organ development by activating a TGR5 receptor for the first time, and a new direction is provided for liver health maintenance and regenerative medicine.

The invention relates to the technical field of biomedical detection, and discloses a solid tumor driving gene detection reading standardization system and kit for cold light enzyme. Comprises: a multi-source data acquisition module configured to acquire cold light enzyme detection data of a plurality of detection platforms; the control module is configured to construct a standardized model based on the large-scale multi-center cooperation data set and process the cold light enzyme detection data by using the standardized model, and comprises the following steps: calling a corresponding standardized sub-model according to sample type information and detection platform parameters corresponding to the cold light enzyme detection data, converting an original fluorescence signal value of the microplate reader into a standardized reading for generating a corrected detection result; and through consistency verification of cross-platform detection data, deviation calibration of a multi-center data set and clinical result correlation analysis, dynamic verification and iterative optimization are carried out on the accuracy of the standardized model. Systematic compensation of detection heterogeneity is realized, and the blank in the field of multi-source data standardization processing is filled.

The invention provides lipid nanoparticles taking calcium phosphate as an inner core and a preparation method of the lipid nanoparticles, and relates to the technical field of biological medicines. The lipid nanoparticles provided by the invention comprise calcium phosphate nanoparticles, DOPA (Dioctyl-Phthalate Polyamide), ionizable cationic lipid, cholesterol and PEG (Polyethylene Glycol) lipid, the calcium phosphate nano-particles are inner cores of the lipid nano-particles. The lipid nanoparticle has excellent biocompatibility and safety and a stable structure, and can improve the stability and delivery efficiency of nucleic acid (such as Luciferase mRNA) as a carrier, so that effective treatment of diseases is realized.

The present disclosure relates in some aspects to methods, systems, and kits for sequencing a template nucleic acid molecule using one or more polymerase-luciferase fusion proteins. In some embodiments, different types of polymerase-luciferase fusion proteins and are sequentially contacted with a template nucleic acid in the presence of different types of nucleotides under conditions that stabilize a ternary complex between a given polymerase-luciferase fusion protein, the template nucleic acid molecule, and a nucleotide when the nucleotide is complementary to the template nucleic acid. By imaging the sample to detect luminescence, the base of the template nucleic acid can be identified.

The present application relates to the technical field of lung adenocarcinoma ferroptosis visualization, in particular to a method for constructing a visual mouse model for real-time monitoring of lung adenocarcinoma ferroptosis process and application. The present application discloses a method for constructing a mouse model carrying exogenous Luciferase (LUC) and eGFP fragments, using live imaging technology to monitor the changes of key markers of ferroptosis in lung adenocarcinoma cells in real time, revealing the dynamic changes of cell signaling network in the process of ferroptosis, and evaluating the influence of different interventions on ferroptosis. The visual mouse model for real-time monitoring of lung adenocarcinoma ferroptosis process provided by the present application can not only deeply explore the sensitivity and resistance mechanism of lung adenocarcinoma cells to ferroptosis, provide theoretical basis and technical support for the development of new therapies based on ferroptosis, but also evaluate the influence of different drugs on ferroptosis, providing a new idea for the treatment of lung adenocarcinoma.

The invention belongs to the technical field of gene therapy, and particularly relates to a compound for regulating INHBE expression and application thereof. According to the present invention, through the siRNA sequence design, the double-chain siRNA sequence RRG008-65 of the target INHBE is screened, and the expression of the INHBE can be knocked down to 29.34% under the concentration of 0.04 nM by using the sequence; on the basis, the siRNA conjugate RRG008-100 and the siRNA conjugate RRG008-112 are obtained after chemical modification and ligand modification are carried out on the siRNA conjugate RRG008-100 and the siRNA conjugate RRG008-112. When the concentration of RRG008-100 is 0.04 nM, the expression of INHBE can be knocked down to 24.06%, and the expression of INHBE can be knocked down to 24.06%. The IC50 of the RRG008-112 for inhibiting INHBE Luciferase is 0.033 nM which is obviously lower than that in the prior art, and meanwhile, a machin experiment proves that the activity and duration of the RRG008-112 for inhibiting the expression of INHBE mRNA in the liver in vivo are superior to those in the prior art.

The invention provides a preparation method and application of an in-vivo CAR-T cell for treating interstitial lung disease, and provides a T cell of a chimeric antigen receptor which is modified by genetic engineering and is used for expressing targeted fibroblast activating protein. A targeted fibroblast activation protein (FAP) CAR-T cell therapy model is constructed by targeting CD5 entrapped mRNA nano-liposome (LNP) transfection and lentiviral vector, and the method comprises the following steps: firstly, verifying the difference of the killing ability of FAP CAR-T constructed by CD5 LNP-mRNA and lentivirus in a 293T cell which stably co-expresses FAP, Luciferase and mCherry, and then verifying the difference of the killing ability of FAP CAR-T constructed by LNP-mRNA and lentivirus in the 293T cell which stably co-expresses FAP, Luciferase and mCherry; further verifying the effectiveness of the CAR-T cells constructed by transfecting the CD5 LNP-mRNA on cell lines of human fibroblasts (CDD19Lu, LL29 and LL97A) and primary fibroblasts of human and mice in vitro, and further verifying the effectiveness and safety of the CD5 LNP-FAP CAR-T in treatment of pulmonary fibrosis through in-vivo experiments in animals. A novel and effective anti-fibrosis treatment thought is provided for patients with fibrosis interstitial lung diseases.

The application discloses application of Neuropathiazol in preparation of a product for treating pancreatic cancer and belongs to the technical field of tumor drugs. The compound is screened through Cell Counting Kit-8, promoter luciferase, cell immunofluorescence (Ki-67, CC3, Map2, Tuj1) staining and other experiments, and is further evaluated by using an immunodeficient mouse transplanted tumor model and a wild-type C57 mouse orthotopic transplanted tumor model. The experimental results show that the compound can not only significantly inhibit the proliferation of in-vitro cultured pancreatic cancer cell strains Panc-1 and SW1990, but also promote the differentiation of tumor cells into neurons, and can effectively inhibit the growth of transplanted tumors in mice. Since the compound is safe, efficient and has an ideal anticancer effect, the compound will be developed into a new type of therapeutic drug for resisting pancreatic cancer, and will certainly have a good application prospect.

The application provides indole diterpene alkaloid compounds and a preparation method and application thereof, and relates to the field of marine drugs. Two novel indole diterpene alkaloid compounds are disclosed, which are named brefeldindole E and brefeldindole F, the structural formulae of which are shown as formula (I) and formula (II) respectively. Both of them have a significant inhibitory effect on LPS-induced NF-kappa B luciferase at a concentration of 10 micromoles, further inhibit RANKL-induced osteoclast precursor cell BMMs cell differentiation into osteoclasts, and have no obvious cytotoxicity. The compounds can effectively replace existing bisphosphonate drugs and denosumab, inhibit the bone resorption activity of osteoclasts, are ideal candidate compounds for developing new osteoclast differentiation inhibitor drugs, and are used for preventing and treating osteoclast-related osteolytic diseases.