12 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.

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 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 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 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.