14899results about "Genetically modified cells" patented technology

Compositions and methods for editing deleterious mutations associated with Alpha-1 Antitrypsin Deficiency (A1AD). In particular embodiments, the invention provides methods for treating A1AD using a modified adenosine base editor with improved on-target editing and decreased off-target editing to correct mutations associated with A1AD.

A recombinant humanized collagen and a preparation method therefor. The recombinant humanized type III collagen can be efficiently and stably expressed in three expression systems of yeast, Escherichia coli, and CHO cells, and has the biological activity superior to that of a commercially available collagen product. The recombinant humanized type III collagen has structural characteristics and functional activity similar to those of a natural collagen, and thus has wide application prospects in biomedical materials, tissue engineering products, cosmetics, foods, health care products or drugs, medical instruments, medical cosmetology and the like.

The invention relates to a nano antibody targeting cadherin 17 and application thereof, and relates to the technical field of immunology and molecular biology, the amino acid sequence of a complementary determining region of the nano antibody targeting cadherin 17 comprises CDR1 shown as any one of SEQ ID NO: 21-32, CDR2 shown as any one of SEQ ID NO: 34-45, and CDR3 shown as any one of SEQ ID NO: 47-58. According to the invention, an alpaca immune phage display library is constructed, a series of nano antibody molecules targeting cadherin 17 (CDH17) are screened from the alpaca immune phage display library, and functions and applications of the nano antibody are verified at various levels of CDH17 antigen protein, CDH17 target cells, CDH17 CAR-T cells and the like.

Disclosed herein are engineered rhizobia having nif clusters that enable the fixation of nitrogen under free-living conditions, as well as ammonium and oxygen tolerant nitrogen fixation under free-living conditions. Also provided are methods for producing nitrogen for consumption by a cereal crop using these engineered rhizobia.

The invention provides recombinant human III-type collagen which is of a triple-helix structure and assembled into collagenous fibers and application of the recombinant human III-type collagen, and belongs to the technical field of bioengineering. Collagen fragments capable of forming a triple helix structure are screened out through a bioinformatics algorithm, the fragments are connected in series, a key C-terminal structural domain and a specific amino acid sequence are combined, and formation of a stable triple helix advanced structure and assembly of collagenous fibers are effectively promoted. The recombinant human III type collagen is efficiently expressed through a eukaryotic expression system, and shows excellent stability, low immunogenicity and good biocompatibility. Experimental results show that the prepared recombinant human III-type collagen not only has a natural triple helix structure, but also can be further assembled into macromolecular collagen fibers, is suitable for multiple fields such as medical beauty, bioengineering and tissue repair, and has wide application prospects and market values.

The present disclosure relates in some aspects to methods, cells, and compositions for preparing cells and compositions for genetic engineering and cell therapy. Provided in some embodiments are streamlined cell preparation methods, e.g., for isolation, processing, incubation, and genetic engineering of cells and populations of cells. Also provided are cells and compositions produced by the methods and methods of their use. The cells can include immune cells, such as T cells, and generally include a plurality of isolated T cell populations or types. In some aspects, the methods are capable of preparing of a plurality of different cell populations for adoptive therapy using fewer steps and / or resources and / or reduced handling compared with other methods.

The disclosure relates to immune cells for treatment of blood cancer comprising an activator receptor comprising an extracellular ligand binding domain specific to an activator antigen expressed by blood cancer cells and an inhibitor receptor comprising an extracellular ligand binding domain specific to an inhibitor antigen expressed by non-cancerous blood cells.

The disclosure provides compositions and methods for the preparation, manufacture and use of an adeno-associated virus (AAV) particle for the vectorized delivery of an antibody molecule that binds to tau.

The invention provides compositions and methods for treating cancer by using immune effector cells (e.g., T cells, NK cells) engineered to conditionally express an agent which enhances the immune effector response of an immune effector cell that expresses a Chimeric Antigen Receptor (CAR). The conditional agents described herein include agents that target a cancer associated antigen, e.g., a CAR, agents that inhibit one or more checkpoint inhibitors of the immune response, and a cytokine.

The invention belongs to the technical field of molecular biology and genetics, and particularly relates to a method for editing cattle genes based on Pro-iCHI. The invention provides a method for editing cattle genes based on Pro-iCHI, which comprises the following steps: transiently expressing protamine in cattle AG-haESCs after gene editing, and injecting into mature oocytes to obtain reconstructed embryos. The protamine can remove abnormal DNA methylation generated by an oocyte cytoplasm injection haploid stem cell technology, cell nucleuses are compressed into a sperm-like structure, the obtained bovine Pro-iCHI embryo can smoothly develop into a blastocyst, and the ratio of the blastocyst to the blastocyst obtained by in-vitro fertilization is equivalent. Moreover, the protamine coding gene is inserted into the saccharomyces cerevisiae protein expression vector and then is subjected to transient expression, so that the protamine coding gene cannot be integrated into a genome to cause insertion of an exogenous gene while abnormal DNA methylation is erased.

The invention provides a method of treating an adult subject having a hematological cancer, comprising administering to the subject selected dosage regimens comprising a plurality of immune effector cells expressing a CAR molecule.

The invention provides an artificial nucleic acid molecule which is used for improving the expression quantity of target amino acid, polypeptide or protein. The artificial nucleic acid molecule at least comprises a target 5'untranslated region (UTR), a target coding region (CDS) and a target 3 'untranslated region (UTR). Wherein the sequence of the target 5 'UTR is one of the following sequences: 5' UTR of a high-expression gene and a 5 'UTR variant of the high-expression gene. The sequence of the target 3 'UTR is one of the following sequences: 3' UTR of a high-expression gene and a 3 'UTR variant of the high-expression gene. Optionally, the artificial nucleic acid molecule may further comprise, for example, a 5 '-end cap structure (Cap), a PolyA tail. The 5 'UTR and the 3' UTR have regulating effects on translation and stability of nucleic acid molecules, so that the 5 'UTR, the 3' UTR and variants thereof are selected from high-expression genes, the nucleic acid molecules can be further stabilized and are not easy to degrade, and the amount of protein or polypeptide obtained by translation of the nucleic acid molecules can be increased. The invention also provides methods for making, delivering, and using such artificial nucleic acid molecules, as well as the use of the artificial nucleic acid molecules for the treatment and / or prevention of related diseases or disorders.

The invention discloses a site for stably expressing protein in a CHO cell gene NW023276806.1 and application of the site, and belongs to the technical field of biological genes. The site belongs to a fixed position in a CHO cell genome, different protein genes are introduced based on a micro-homologous end connection mechanism through a CRISPR / Cas9 tool, and stable expression is carried out. By adopting a site-specific integration method, a target gene is integrated to a stable expression area in a site-specific manner, repeated high-expression monoclonal screening is effectively avoided, and an MMEJ mechanism is introduced to integrate a donor fragment, so that the research and development time for constructing a stable expression cell strain in biological pharmacy can be effectively shortened, and the cost is reduced.

The present invention provides a method for packaging a BaEV retroviral vector. The method includes constructing a stable BaEV retroviral packaging cell line, transfecting a retroviral vector plasmid containing a target sequence into a virus-producing cell line, where the virus-producing cell line is a HEK293T cell line or its derivative cell line, to harvest the transiently transfected and virus-producing retroviral vector, mixing it with the stable BaEV retroviral packaging cell line, and promoting the transduction of the retroviral vector into the stable BaEV retroviral packaging cell line by horizontal centrifugation. The present invention can achieve the packaging of a BAEV-type retroviral vector carrying a CD19CAR foreign gene plasmid, construct a BAEV-type retroviral vector-producing cell line, and produce a high-titer BAEV-type retroviral vector.

The invention discloses an anti-interleukin-6 antibody or a reagent and a kit for detecting interleukin-6, and relates to the field of antibodies. The anti-interleukin-6 antibody disclosed by the invention comprises a heavy chain complementarity determining region and a light chain complementarity determining region, provides an important raw material source for detection of interleukin-6, and has good affinity or activity.

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 discloses a method for improving beef quality, an action mechanism and an experimental method, in a cattle body, vitamin A can activate the expression of EBF2 through an active metabolite RA of the vitamin A, and the EBF2 can inhibit the transcription process of CYP26B1 in a targeted manner to maintain the activity of a retinol signal channel, so that PPAR gamma and downstream lipid metabolism related genes thereof are activated, and the activity of the retinol signal channel is improved. Fatty acid transport and lipid accumulation in fat cells in the cattle muscle are promoted, fat deposition in the cattle muscle is promoted, and the beef quality is improved. By constructing the molecular network for regulating and controlling the formation of the fat in the cattle muscle, a complex regulation and control mechanism for controlling the fat deposition in the cattle muscle can be understood more deeply, a method for improving the beef quality based on regulating and controlling the vitamin A mediated key factor EBF2 is provided, and a new way is provided for improving the beef quality in a targeted manner.

The invention provides a porcine epidemic diarrhea virus S protein mutant, a nucleic acid molecule, a subunit vaccine and application. Compared with wild type porcine epidemic diarrhea virus S protein, the porcine epidemic diarrhea virus S protein mutant has the advantage that amino acid at the 1076th site, amino acid at the 1077th site, amino acid at the 1016th site and amino acid at the 1017th site are mutated into proline. The natural trimer which is stable and maintains the conformation of the S protein before fusion is constructed, the safe and efficient PEDV subunit vaccine is obtained, and the use effect and immunogenicity of the porcine epidemic diarrhea vaccine can be improved; moreover, compared with a wild type S protein, the porcine epidemic diarrhea virus S protein mutant disclosed by the invention has a higher expression quantity.

The invention discloses an anti-cortisol antibody or a cortisol detection reagent and kit, and relates to the field of antibodies. The anti-cortisol antibody disclosed by the invention comprises a heavy chain complementarity determining region and a light chain complementarity determining region, provides an important raw material source for detection of cortisol, and has good affinity or activity.

The invention discloses a CA9-targeted chimeric antigen receptor and application thereof, and belongs to the technical field of biological medicines. The CA9 targeting chimeric antigen receptor provided by the invention comprises a CD8 alpha leading signal peptide, an anti-CA9 single-chain variable fragment, a CD8 alpha hinge, a CD8 transmembrane structural domain and a CD3 zeta intracellular costimulatory signal transduction structural domain. According to the invention, an anti-CA9 single-chain variable fragment is used as an antigen binding structural domain to construct a chimeric antigen receptor (CAR) molecule, the macrophage expressing the anti-CA9CAR can realize specific targeting and phagocytosis killing of CA9 positive kidney cancer cells, and the constructed lipid nanoparticle delivery system can effectively transfer a gene expression vector, so that the gene expression vector can realize effective target site delivery, and the CA9 positive kidney cancer cells can be effectively targeted and killed. The constructed hydrogel-liposome combined drug delivery system has the performance of relieving toxic and side effects, controllable release and hemostasis.

The present disclosure provides bispecific chimeric antigen receptors targeting CD20 and BCMA. The CAR may comprise an scFv targeting CD20 and an scFv targeting BCMA, a hinge region, a transmembrane domain, a co-stimulatory region, and a cytoplasmic signaling domain. The chimeric antigen receptors can be used to treat autoimmune disorders or cancer.

The invention belongs to the technical field of biology, and provides a nanometer antibody targeting Nectin-4, and a nucleic acid molecule and an expression vector for coding the nanometer antibody targeting Nectin-4. The invention also provides a chimeric antigen receptor which comprises an extracellular antigen binding domain, a transmembrane domain and an intracellular domain, and the extracellular antigen binding domain is a Nectin-4 nano antibody; the invention also provides a chimeric antigen receptor CAR which contains the nano antibody as scFv and co-expresses CXCR1, a CAR-T cell constructed by the chimeric antigen receptor CAR, and an enhanced CAR-T cell obtained by modifying the CAR-T cell with microbial short-chain fatty acid, so as to improve the infiltration and anti-tumor activity of the CAR-T cell targeting Nectin-4.

The invention relates to the technical field of recombinant protein expression, in particular to recombinant human-derived III-type collagen as well as a preparation method and application thereof. The invention provides a recombinant human source III type collagen. The amino acid sequence of the recombinant human source III type collagen is shown as SEQ ID NO. 1. The recombinant human-derived III-type collagen prepared by the invention can be better applied to a human body, better exerts biological functions of the collagen, avoids repulsive interaction, is high in safety, can effectively promote cell proliferation and oxidation resistance, and is high in expression yield, stable in batches and good in quality; the recombinant human III type collagen prepared by the invention has the potential of being applied to preparation and production of biological medicines and cosmetics.

The invention discloses a nanometer antibody targeting FAP, a CAR-T cell and application of the nanometer antibody and the CAR-T cell. The invention provides nano-antibodies VHH1 and VHH2 capable of simultaneously recognizing human or mouse FAP. The nano-antibodies VHH1 and VHH2 are used as antigen binding regions to be constructed in a human or mouse CAR structure, and human or mouse FAP-CAR-T cells with a killing function can be prepared. Results of the mouse FAP-CAR-T cell or the human FAP-CAR-T cell provided by the invention show that the FAP-CAR-T cell has a good treatment effect on removal of FAP positive cells, and is a novel FAP-CAR-T cell which is expected to be applied to clinical solid tumor CAR-T therapy.

The invention relates to a humanized nano antibody targeting cadherin 17 and application of the humanized nano antibody, and relates to the technical field of immunology and molecular biology. The complementarity determining region of the humanized nano antibody comprises a CDR1 of which the amino acid sequence is shown as SEQ ID NO: 2, a CDR2 of which the amino acid sequence is shown as SEQ ID NO: 4 and a CDR3 of which the amino acid sequence is shown as SEQ ID NO: 6; a framework region of the humanized nano antibody comprises an FR1 of which the amino acid sequence is shown as any one of SEQ ID NO: 1 and SEQ ID NO: 8, an FR2 of which the amino acid sequence is shown as any one of SEQ ID NO: 3, SEQ ID NO: 9 and SEQ ID NO: 11, an FR3 of which the amino acid sequence is shown as any one of SEQ ID NO: 5 and SEQ ID NO: 10, and an FR4 of which the amino acid sequence is shown as SEQ ID NO: 7. According to the humanized nano-antibody, the immunogenicity of the nano-antibody is reduced, the targeted binding activity and killing activity are improved, and the humanized nano-antibody can be used for developing immunodetection reagents, CAR-T / NK cell drugs and antibody drugs.

The invention relates to a drug delivery carrier based on extracellular vesicles and application thereof.The drug delivery carrier contains extracellular vesicle scaffold protein and recombinant membrane fusion protein, the recombinant membrane fusion protein comprises a first fragment and a second fragment, the amino acid sequence of the first fragment is shown as SEQ ID NO: 5, and the amino acid sequence of the second fragment is shown as SEQ ID NO: 6; the second fragment has n amino acids, the sequence of the second fragment is the sequence of the first n amino acids at the N end of the amino acid sequence as shown in SEQ ID NO: 7, and n is an integer between 50 and 200. According to the drug delivery carrier disclosed by the invention, the efficiency of releasing drug active molecules before reaching lysosome is remarkably improved, and the drug effect of extracellular vesicle drugs can be improved.

The invention relates to the technical field of biological medicine, and provides a targeting exosome capable of delivering nucleic acid drugs and a preparation method of the targeting exosome. The exosome comprises a fusion polypeptide, the fusion polypeptide comprises a targeting protein molecule, an exosome transmembrane protein and a nucleic acid binding domain, and the exosome can load a nucleic acid molecule through binding of the nucleic acid binding domain. According to the invention, engineering modification is carried out on the exosome, so that nucleic acid molecules can be efficiently loaded into the exosome, and meanwhile, the exosome is endowed with the capability of targeting to specific target cells, tissues and organs.

The invention provides a specific T cell receptor targeting KRAS G12V mutant epitope peptide (such as an amino acid sequence as shown in SEQ ID NO: 2) and anti-tumor application of the specific T cell receptor. The specific T cell receptor is composed of two peptide chains alpha and beta and can be specifically combined with a VVGAVGVGK-HLA-A * 11: 01 compound. The invention also provides an antigen binding fragment of the specific T cell receptor, a nucleic acid sequence for coding the T cell receptor, a vector containing the nucleic acid sequence, an engineered cell for expressing the T cell receptor, a composition containing the T cell receptor and application thereof. In addition, the invention also provides a method for screening the KRAS G12V specific T cell receptor. The specific T cell receptor and the antigen binding fragment thereof provided by the invention can be used as an immune effect activator to stimulate the immune response of a body, so that the effect of resisting diseases such as tumors and the like is achieved.

The invention discloses a MAGE-A10 specific TCR (T cell receptor) and an application thereof. The invention provides a TCR (T cell receptor) which has high antigen sensitivity and can remarkably kill target cells, and also provides nucleic acid for coding the TCR, a carrier comprising the nucleic acid, a cell, a pharmaceutical composition and application of the TCR in preparation of the pharmaceutical composition for treating tumors and screening of antigen detection products.