598 results about "Transfection" patented technology

Recombinant NK cells, and especially recombinant NK-92 cells express a chimeric antigen receptor (CAR) having an intracellular domain of FcεRIγ. Notably, CAR constructs with an intracellular domain of FcεRIγ had a substantially prolonged duration of expression and significantly extended cytotoxicity over time. The CAR may be expressed from RNA and DNA, preferably as a tricistronic construct that further encodes CD16 and a cytokine to confer autocrine growth support. Advantageously, such constructs also enable high levels of transfection and expression of the recombinant proteins and provide a convenient selection marker to facilitate rapid production of recombinant NK / NK-92 cells.

The invention relates to the technical field of cytology, in particular to a grass carp brain astrocyte line and application thereof, the grass carp brain astrocyte line is preserved in China Center for Type Culture Collection on May 7, 2025, and the preservation number of the grass carp brain astrocyte line is CCTCC NO: C2025149. The grass carp brain astroglia cell line provided by the invention has the capability of efficiently proliferating GCRV-II, and the virus titer of the GCRV-II replicated in the cell line at least can reach 1.38 * 10 < 8 > pfu / mL or above; after the GCRV-II is blindly passed for 5 generations in a grass carp astroglia cell line, the grass carp can still have typical bleeding symptoms and death due to the virus. And the exogenous plasmid transfected grass carp brain astroglia cell line has similar transfection efficiency to commercial grass carp kidney cells. Therefore, the invention lays an important foundation for deep research of pathogenic mechanism of GCRV-II, vaccine preparation, antiviral drug screening and prevention and control of grass carp viral hemorrhagic disease.

The invention discloses a self-replicating RNA (Ribonucleic Acid) molecule, a porcine rotavirus RNA vaccine and application thereof, and relates to the technical field of vaccines. According to the present invention, the coding sequence of the porcine rotavirus G9P23 type VP4 protein is optimized, such that the fact that the saRNA prepared by using the nucleotide sequence represented by the SEQ ID NO: 10 can secrete the high porcine rotavirus G9P23 type VP4 protein antigen after the cell transfection is found; after the optimized porcine rotavirus G9P23 type VP4 protein is used for immunizing experimental animals, the level of neutralizing antibodies generated by the animals can be improved, and the challenge protection effect is enhanced. Therefore, the nucleic acid molecule provided by the invention can be used for developing drugs and vaccines for causing protective immune response in experimental animal bodies. Compared with a traditional vaccine, the vaccine provided by the invention has good immunogenicity and safety, and has a good application prospect.

The invention relates to the technical field of gene engineering. The invention provides a functional sequence shRNA1 (short hairpin ribonucleic acid 1) for silencing a circular RNAh-hsacirc005838, and further provides a lentiviral vector of the functional sequence shRNA1 and a construction method of the lentiviral vector. The invention also provides a lentivirus packaging titer kit containing the lentiviral vector with the functional sequence shRNA1, and also provides an application of the lentiviral vector with the functional sequence shRNA1 or the lentiviral vector with the functional sequence shRNA1 in preparation of a kit for breast cancer diagnosis. According to the invention, an interference target sequence siRNA1 is obtained by aiming at the annular target gene RNA h-hsacirc005838, then the finally obtained functional sequence shRNA1 can directly and efficiently infect cells and silence the annular RNAh-hsacirc005838 sequence, and the transfection effect is stable.

The invention discloses application of arabinogalactan protein family genes PbrAGP16 in promoting growth of pear pollen tubes, and belongs to the technical field of plant genetic engineering. According to the invention, the gene PbrAGP16 is cloned from pollen of Dangshan pears by using a plant gene cloning technology, and the nucleotide sequence of the gene PbrAGP16 is shown as SEQ ID No.1; the amino acid sequence coded by the gene is SEQ ID No.2 in a sequence table. Researches show that PbrAGP16 can promote growth of pear pollen tubes, the change of PbrAGP16 expression is related to the change of the content of cellulose at the top ends of the pollen tubes, and PbrAGP16 also participates in a regulation mechanism of a self-incompatible reaction. The pollen magnetic transfection technology is adopted to research the function of the gene in the pollen tube, and a wide application prospect is provided for improving the pollination efficiency.

The present disclosure provides for improved compositions of ionizable lipid nanoparticles for the delivery of therapeutic nucleic acids to cells. Anionic phospholipids, including phosphatidylserine and phosphatidylglycerol are included in the lipid nanoparticles to increase the transfection efficiency in human dendritic cells. The further incorporation of mono-unsaturated alkyl chain analogs in dimethylaminopropyl-dioxolane or heterocyclic ketal ionizable lipids in the formulation demonstrated high levels of transfection in human dendritic cells, compared to other ionizable lipids in the same family, and demonstrated good stability to oxidative damage. Finally, the use of an ammonium salt of phosphatidylserine allows for the efficient production of PS-targeted LNPs.

The invention discloses an application of a compound in preparation of a product for enhancing the metabolic activity of hematopoietic stem cells and treating metachromatic white matter dystrophy. The compound comprises Azoramide. Azoramide is adopted to treat transfected hematopoietic stem cells, and cell apoptosis, cell necrosis and mitochondrial respiration conditions of the hematopoietic stem cells are obviously improved, so that the hematopoietic stem cells are expected to be applied to enhancement of transfection efficiency and metabolic activity of the hematopoietic stem cells transfected by the ARSA gene and reduction of ROS generation, and the effectiveness of the hematopoietic stem cells in treatment of MLD diseases is enhanced.

Provided in the present invention is a nitrogen-containing heterocycle-branched cationic lipid. The structure thereof is as represented by general formula (1), wherein the definition of each symbol is consistent with that described herein. The nitrogen-containing heterocycle-branched cationic lipid has a nitrogen-containing heterocycle as a branched core, an amino acid or an amino acid derivative as a linker arm, and a saturated or unsaturated hydrocarbon group as a tail chain; and has the advantages of low toxicity, low immunogenicity and high biocompatibility. In the new cationic lipid of the present invention, the electropositivity of the nitrogen on the nitrogen-containing heterocycle is delocalized to the whole ring, so that the cationic lipid shows higher transfection efficiency and lower cytotoxicity. The new cationic lipid of the present invention contains one or more biodegradable groups between the nitrogen-containing heterocycle and the hydrophobic tail, and the presence of the degradable groups enables a lipid nanoparticle (LNP) prepared from the cationic lipid to be timely degraded in the endosome, such that the problem in the prior art whereby LNPs prepared from a non-degradable lipid accumulate in the endosome and acidify the endosomal environment, thereby hindering the endosomal escape of a drug molecule (such as a nucleic acid) and thus the drug delivered into a cell not being able to function is solved.

The invention belongs to the technical field of biological medicine, and particularly relates to an engineered cell microvesicle and a delivery system based on the engineered cell microvesicle, the system realizes efficient preparation of 1-5 [mu] m cell microvesicles, and the cell microvesicles have a large space volume and can be used for preparing the cell microvesicles. The carrier can be used for loading and delivery of target protein, polypeptide and recombinase which are specifically expressed in mother cells. The system transfects mother cells through lentivirus transfection or plasmid transfection to further produce cell microvesicles, and the microvesicles can load more goods and inherit membrane proteins of the mother cells, and can also effectively load intracellular proteins to realize effective delivery. By virtue of good structural stability, high immunogenicity and excellent biocompatibility, the cell microvesicle reduces systematic toxic and side effects of a traditional carrier, is expected to become an effective drug delivery system, and has great application potential in the field of gene therapy.

The invention provides a stem cell exosome for overexpressing A beta degrading enzyme as well as a preparation method and application of the stem cell exosome. Specifically, the invention provides an application of the stem cell exosome overexpressing A beta degrading enzyme in preparation of drugs, and the drugs are used for at least one of the following: prevention, improvement or treatment of cognitive impairment related diseases; neuronal cell damage is improved; wherein the stem cells for overexpressing the A beta degrading enzyme are prepared through lentivirus transfection. According to the stem cell exosome overexpressing the A beta degrading enzyme, the content of the A beta degrading enzyme is obviously higher than that of the exosome prepared by a traditional method. Therefore, when the compound is prepared into a medicine for treating or preventing cognitive impairment related diseases, the medicine can be efficiently conveyed to the brain, and the concentration of the medicine in the brain is remarkably improved, so that the curative effect is enhanced, the dosage of the medicine can be reduced, the side effect is reduced, and the prognosis of brain diseases is effectively improved.

The present invention belongs to the field of drug delivery. Provided is an asymmetric cationic lipid containing a branched piperazine ring and having a structure as shown in general formula (1), wherein the definition of each symbol is consistent with that described in the text. In the cationic lipid of the present invention, a degradable group is introduced at an appropriate position, and the presence of the degradable group makes an LNP-pharmaceutical composition prepared therefrom be degraded at an appropriate time in vivo and have a low cytotoxicity, which solves the problems in the prior art that an LNP-pharmaceutical composition prepared from a non-degradable lipid can accumulate in endosomes and acidify the endosomal environment, so that the endosomal escape of a drug (e.g., mRNA) is hindered, and the drug delivered into cells cannot fully function. In the cationic lipid of the present invention, a heterofunctionalized small molecule containing a piperazine ring is used as a raw material. The preparation process is simple and convenient, has a lower cost, and is more environmentally friendly. The LNP-mRNA composition prepared therefrom has the advantages of low toxicity, high biocompatibility, high cell transfection, etc.

The invention relates to the technical field of biological medicine. The invention provides highly branched poly (beta-amino ester) and a preparation method thereof. The invention also provides lipid nanoparticles which contain the compound and are used for efficiently delivering mRNA, a preparation method of the lipid nanoparticles, and application of the lipid nanoparticles in preparation of CD5 antibody targeted nucleic acid drug delivery carriers. The highly branched poly (beta-amino ester) prepared by the scheme of the invention has better mRNA binding capacity and in-vivo stability, the obtained lipid nanoparticles have high transfection efficiency, and efficient transfection of specific tissues or cells can be better realized.

The invention discloses a nucleic acid transfection system and method based on automatic control, and relates to the technical field of genetic engineering.The method comprises the steps that after target cells are intelligently cultured to be in a suitable state, an automatic system selects a transfection reagent and prepares a compound according to cell types; the cells and the compound are mixed through low shear force and then incubated; multi-modal monitoring equipment is used for tracking nucleic acid distribution and cell states in real time, and incubation conditions are dynamically adjusted; analyzing the monitoring data based on a machine learning algorithm and optimizing transfection parameters; after transfection, culture and multi-dimensional analysis are automatically executed. The system correspondingly comprises a cell culture module, a reagent preparation module, a mixed incubation module, a real-time monitoring module, a dynamic optimization module and a subsequent analysis module. Through closed-loop automatic control and intelligent optimization, the transfection efficiency and stability are remarkably improved, the cytotoxicity is reduced, manual intervention is reduced, and a standardized solution is provided for recombinant gene expression.

The present disclosure provides for improved compositions of ionizable lipid nanoparticles for the delivery of therapeutic nucleic acids to cells. Cationic ionizable lipids are engineered with improved stability to oxidative degradation while in storage, while retaining high transfection activity or potency in cells. These lipids are designed to be biodegradable, thus improving the tolerability of nanoparticles formed with them in vivo. In addition, targeting of these nanoparticles in a highly specific manner to dendritic cells is provided for through inclusion of antibody conjugates directed against cell surface receptors.

The invention relates to the field of biological medicine, in particular to a cationic lipid compound capable of being used for nucleic acid delivery and a preparation method and application thereof. The LNP prepared from the cationic lipid compound with the brand new structure has the advantages of high transfection efficiency and the like, and is particularly only distributed at an injection part.

The preparation and evaluation method comprises the following steps: 1, extracting a heavy-chain DNA fragment and a light-chain DNA fragment, carrying out recombinant transformation by using an overlapping PCR method to construct an scFv gene, 2, constructing an expression vector, culturing the expression vector, carrying out sequencing identification to obtain an scFv bacterial solution, recombining the scFv bacterial solution with a mouse Fc fragment, and identifying a sequence to obtain an MN8 bacterial solution, the method comprises the following steps: 1, preparing an MN8 bacterial liquid, 2, extracting the MN8 bacterial liquid, 3, carrying out inoculated culture on the MN8 bacterial liquid to obtain an MN8 genetically engineered antibody bacterial liquid, 4, extracting MN8 genetically engineered antibody bacterial liquid plasmids and carrying out transfection expression purification, and 5, carrying out antibody titer and thermal stability determination on the MN8 genetically engineered antibody. The scFv single-chain antibody obtained by recombination according to the method has the advantages of small molecular weight, high penetrability and the like, the stability and the sensitivity of an immunoassay method of the scolosporins can be improved, and a certain basis is provided for rapid determination of the scolosporins.

The present disclosure provides a method for preparing natural killer (NK) cells with increased secretion of cytokines and increased cytotoxicity. The present invention uses IL-12 during transfection to generate CAR-NK cells with increased functional activities (cytotoxicity and secretion of cytokines) against tumor cells.

Provided herein are formulations for oral delivery or administration of therapeutic nucleic acids. An oral formulation can include: a nucleic acid; at least one casein protein; and a chitosan. In some embodiments, the formulations are liposome-free formulations having no cationic lipids typically used as transfection reagents in conventional nucleic acid formulations for oral delivery. The formulations provided herein find use in treating a condition associated with inflammation and / or fibrosis, or a cardiometabolic disorder by oral administration.

The present invention belongs to the field of drug delivery. Provided is an asymmetric cationic lipid containing multiple tertiary amines, of which the structure is shown as a general formula (1), wherein the definition of each symbol is consistent with that in the description. In the present invention, a degradable group is introduced into a proper position of the cationic lipid, and the presence of the degradable group enables an LNP-drug composition prepared from the cationic lipid to be degradable in endosomes at a proper time and to have small cytotoxicity, thereby solving the problem that in the prior art, LNP-drug compositions prepared from non-degradable lipids will accumulate in endosomes and acidify the endosomal environment, which hinders the endosomal escape of drugs (such as mRNAs) and causes that the drugs delivered into cells fail to sufficiently exert the therapeutic effects. The cationic lipid in the present invention uses a heterofunctionalized piperazine-containing small molecule as a raw material, involves a simple preparation process and a lower cost, and is more green and environmentally-friendly. An LNP-mRNA composition prepared from the cationic lipid has the advantages of low toxicity, high biocompatibility, high cell transfection, etc.

An ionizable cationic lipid compound having a structure as represented by formula (I), which can be used for preparing a lipid nanoparticle (LNP) for the delivery of a therapeutic agent and / or a prophylactic agent. The LNP prepared with the ionizable cationic lipid compound has a better stability and transfection efficiency, and can efficiently and stably deliver a bioactive substance (comprising a nucleic acid, e.g., mRNA) to a target cell or organ, thereby eliciting a highly specific antibody response in vivo.

The invention discloses a nucleic acid tetrahedron nanostructure as well as a preparation method and application thereof. The nucleic acid tetrahedron nanostructure is composed of circRNA for coding an antigen, an ISD chain and two specific single-stranded DNA sequences, is simple and easy to prepare, can enter cells through cell membrane microfossa mediated endocytosis without the help of a transfection reagent, and is positioned to lysosome. The nanostructure is split to release circRNA coding antigen protein, and the circRNA coding antigen protein is delivered to APC and ISD chains in drainage lymph nodes (dLN) to activate cGAS-STING pathways, so that antigen presentation and immune activation are synergistically enhanced, T cells are efficiently and continuously activated, and the effect of tumor immunotherapy is achieved.

The invention belongs to the field of biological medicine, and provides circular RNA for treating multiple endocrine tumor type I and application of the circular RNA. According to the invention, after different optimization and design are carried out on the circular RNA for coding Menin, the circular RNA with relatively good expression level and long half-life period is finally obtained, and the anti-tumor effect is mainly achieved by supplementing cell-deficient Menin. Compared with the prior art, the circular RNA for expressing the Menin protein provided by the invention is more stable in intracellular expression and longer in half-life period. In an immortalized parathyroid gland cell model, the Menin protein can be expressed by transfecting the circular RNA for 6 hours, and the circular RNA has good treatment potential for multiple endocrine tumor type I.

The invention provides an amino acid skeleton ionizable lipid as well as a preparation method and application thereof, the amino acid skeleton ionizable lipid is modified by taking amino acid as a core, has more ester groups and peptide bonds, and can be quickly hydrolyzed by enzyme after RNA is effectively released in vivo; the transfection body is provided with four tail structures, the cross sectional area of the lipid tail can be increased, drugs such as RNA are helped to escape from an endosome, and then the transfection effect is enhanced; the charge capable of ionizing the lipid is electrically neutral under physiological conditions, so that the cytotoxicity caused by excessive positive charges is reduced, the stability of the lipid nanoparticles is further improved, the cycle time of the loaded nucleic acid medicine is prolonged, and the pharmacokinetic characteristics are improved. The LNP prepared from the ionizable lipid, auxiliary phospholipid, cholesterol and PEG lipid provided by the invention has more excellent nucleic acid carrier performance, and can effectively deliver nucleic acid drugs such as siRNA, mRNA, pDNA and the like into cells to play a role.

The invention belongs to the technical field of biological medicine, and particularly relates to a regulatory element or a combination thereof. The regulatory element comprises a DNA regulatory element and / or an RNA regulatory element; the DNA regulation element is specifically combined with a protein coded by a target gene so as to inhibit transcription of the target gene; a transcription product of the RNA regulation element is specifically combined with a protein coded by a target gene so as to inhibit translation of the target gene, and when the content of the protein coded by the target gene is high, the expression quantity of the protein can be reduced; when the content of the protein coded by the target gene is low, the expression quantity of the protein can be up-regulated, and the decline level of the protein can be reduced, so that the expression level of the target gene is regulated along with the content of the protein, feedback type self-regulation is formed, and the occurrence of overexpression can be reduced under the condition that a carrier containing the target gene and a regulation element is excessively transfected; therefore, the gene is used for preventing and treating diseases caused by insufficient expression of the target gene, and side effects of overexpression of the target gene are reduced while disease symptoms are improved.

The invention discloses a porcine reproductive and respiratory syndrome virus RNA vaccine and application thereof, and relates to the technical field of veterinary biological products. According to the present invention, the coding sequence of the porcine reproductive and respiratory syndrome virus GP5 protein is optimized, and the GP5 protein is connected with the M protein through the connecting peptide, such that the saRNA prepared by using the nucleic acid molecule can produce the efficient expression on the GP5 protein and the M protein after the cell transfection. After the optimized porcine reproductive and respiratory syndrome GP5 protein and M protein fusion protein is immunized, the level of neutralizing antibodies generated by animals can be improved, and the effect of preventing the porcine reproductive and respiratory syndrome virus is achieved. Therefore, the nucleic acid molecule provided by the invention can be used for developing or preparing nucleic acid vaccines. Compared with a traditional vaccine, the vaccine provided by the invention has good immunogenicity and safety, and has a good application prospect.

Provided herein are RNA agent-loaded platelets, methods of preparing RNA agent-loaded platelets, and methods of using RNA agent-loaded platelets. In some embodiments, methods of loading RNA agents into platelets include treating platelets with a RNA agent, a cationic transfection reagent, and a loading buffer that can include a salt, a base, a loading agent, and optionally at least one organic solvent.

The invention relates to the technical field of insulin resistance, in particular to an insulin resistance method for promoting a signal channel based on CTRP9. The method comprises the following steps: constructing a human hepatocyte line plasmid; carrying out cell culture and transfection; detecting the influence of CTRP9 on LO human hepatocyte glucose uptake, PGC-1a gene expression and the phosphorylation level of insulin signal channel related protein; detecting the influence of CTRP9 on glucose uptake of an LO human hepatocyte insulin resistance model, PGC-1a gene expression and the phosphorylation level of insulin signal channel related protein; and carrying out SirT1 / PGC-1a protein expression detection. According to the insulin resistance method for promoting the signal channel based on the CTRP9, a human hepatocyte line is taken as an in-vitro research object, an in-vivo research environment is simulated, a corresponding insulin resistance model is established, and the insulin resistance can be obtained by using technologies such as flow cytometry analysis, RT-PCR (Reverse Transcription-Polymerase Chain Reaction), western blotting and signal channel inhibition. And the treatment effect of CTRP9 on hepatic cell insulin resistance and the specific action mechanism are researched.

An ionizable cationic lipid compound having a structure represented by formula (I), which can be used to prepare lipid nanoparticles (LNP) for the delivery of a therapeutic and / or prophylactic agent. The LNP prepared using the ionizable cationic lipid compound has good stability and transfection efficiency, and can efficiently and stably deliver bioactive substances (including nucleic acids, such as mRNA) to target cells or organs, thereby eliciting a high-specificity antibody response in vivo.

The invention provides a transfection reagent based on blank lipid nanoparticles as well as a preparation method and application of the transfection reagent, and belongs to the technical field of biological medicines. The transfection reagent based on the blank lipid nanoparticles comprises the following components: (1) the blank lipid nanoparticles; (2) a biologically active ingredient; the blank lipid nanoparticles comprise ionizable lipid, phospholipid, cholesterol and polyethylene glycol conjugated lipid. The preparation process of the blank lipid nanoparticles and the transfection reagent is simple, and the preparation can be completed without the help of equipment. The transfection reagent based on the blank lipid nanoparticles is prepared from the blank lipid nanoparticles and nucleic acid, the dosage can be flexibly adjusted according to the requirements of a user, the transfection effect is stable, various cells (such as 293T, Hela, HepG2, TWO3 and MCF7) can be transfected, the cytotoxicity is low, and the transfection effect is good.

The invention relates to an shRNA (short hairpin Ribonucleic Acid) interference sequence of a targeted silence PCSK9 gene and a construction method and lipid-lowering application of a recombinant adeno-associated virus vector of the shRNA interference sequence. Hyperlipidaemia is a metabolic disease characterized by abnormal rising of cholesterol and triglyceride levels in blood, and the design of lipid-lowering drugs is the focus of attention to improvement of hyperlipidaemia. Proprotein convertase subtilisin / kexin type 9 (PCSK9) can be combined with a low-density lipoprotein receptor (LDL-R) and degrade the LDL-R, so that accumulation of LDL-C in blood is further promoted, and hyperlipidemia is caused. Aiming at the key target PCSK9, a specific shRNA interference sequence is designed, and a recombinant adeno-associated virus vector (rAAV) carrying the sequence is constructed by an enzyme digestion-connection method. In-vitro experiments prove that the vector can remarkably reduce the expression level of PCSK9 protein, so that the cyclic utilization of a low-density lipoprotein receptor (LDL-R) is promoted, and the concentration of low-density lipoprotein cholesterol (LDL-C) in plasma is reduced. The rAAV vector provided by the invention has the characteristics of low production cost, high transfection efficiency, lasting action time and the like, and provides a new thought for gene therapy of hyperlipidemia.