16773results about "Foreign genetic material cells" patented technology

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 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 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 belongs to the technical field of cell cryopreservation, and relates to a cryopreservation method based on ultrasonic ice planting. In the method, the nucleation temperature of ice planting is less than-9 DEG C, and the cooling rate from nucleation to icing is less than 5 DEG C / min. Compared with the prior art, the method has the advantages that the time required for cryopreservation of the cells is remarkably shortened, meanwhile, damage to the cells caused by ice crystals generated in the cooling process is reduced, and the problem that an existing cell cryopreservation cooling method cannot meet the nucleation and freezing uniformity of a cell solution in the cooling process is solved.

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 present invention belongs to the technical field of bioengineering. Provided are an IscB mutein and the use thereof. Compared to the wild-type IscB protein, the IscB mutein comprises arginine at at least one of the following amino acid positions: position 84, position 96, position 102, position 111, position 159, position 368 and position 386; preferably, the IscB mutein further comprises arginine at position 401 and position 456. The relatively strong interaction between the IscB mutein and a double-stranded DNA target improves the editing activity for mammalian cell lines, which can greatly enhance the application potential of the IscB mutein as a new-generation underlying tool for gene editing, and promote the use of the IscB mutein in the aspects of precision medicine, animal model construction and crop breeding.

An objective of the present disclosure is to provide anti-CD137 antigen-binding molecules which have immunocyte-activating effect, cytotoxic activity, or anti-tumor activity, and meanwhile have reduced effect on non-tumor tissues such as normal tissues and produce less side effects, and methods of using the same.Anti-CD137 antigen-binding molecules which have immunocyte-activating effect, cytotoxic activity, or anti-tumor activity, and meanwhile have reduced effect on non-tumor tissues such as normal tissues and produce less side effects, are provided by discovering and producing CD137 antigen-binding molecules whose binding activity to CD137 depends on various substances (for example, small molecule compounds) in target tissues. Methods of using the same, pharmaceutical formulations, and such are also provided.The present disclosure also provides an antigen-binding molecule whose binding activity to an antigen varies depending on a small molecule compound, a preparation method thereof, and uses thereof.

The present application relates to the field of biomedicine, and in particular to a tumor antigen peptide and a use thereof. Provided is a tumor antigen peptide, comprising a peptide segment having an amino acid sequence as shown in SEQ ID NO: 1. The provided tumor antigen peptide can be presented by the HLA subtype (HLA-A*11:01) having a high proportion of Chinese population. Additionally, by means of in vitro stimulation, the tumor antigen peptide is verified to effectively stimulate specific T cells, and a specific TCR element is successfully obtained. The present application lays a direct foundation for further developing TCR-T therapy targeting human TP53 R248Q and immunotherapy such as therapeutic vaccines.

The present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to human cMET. The disclosure also provides polynucleotides encoding the antibodies or antigen binding fragments thereof, vectors comprising the polynucleotides, host cells comprising the vectors, methods of generating the antibodies, and compositions comprising the antibodies.

The invention provides a UTR (Untranslated Region) sequence for improving mRNA (Messenger Ribonucleic Acid) stability and translation efficiency and application of the UTR sequence, and particularly provides an mRNA molecule which comprises a coding region for coding polypeptide or protein and a fragment thereof, a 5 'UTR sequence positioned at the upstream of the coding region and / or a 3' UTR sequence positioned at the downstream of the coding region, the 5 'UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 1, and the 3' UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 2; and / or, the 5 'UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 3, and the 3' UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 4. According to the present invention, the mRNA stability and the translation efficiency are improved through the new and optimized UTR sequence, and the UTR sequence can be used as the element for enhancing the RNA expression efficiency in the nucleic acid treatment drug or the mRNA vaccine so as to provide more and better choices for the mRNA therapy.

The invention discloses a grass carp gamma interferon fusion protein as well as a mutant, a coding gene and application thereof. In order to improve the expression quantity or expression efficiency of the grass carp gamma interferon, codon optimization and mutation are carried out on the coding gene of the grass carp gamma interferon to obtain the grass carp gamma interferon mutant. The fusion protein is further obtained by connecting the grass carp gamma interferon mutant with the C end of the grass carp ferritin heavy chain subunit of which the last 17 amino acids are removed through a connecting peptide. In order to further improve the antiviral activity of the fusion protein, the obtained fusion protein is subjected to single-point mutation, and the titer of the fusion protein is remarkably improved. The fusion protein is expressed by a silkworm cell eukaryotic expression system, and the fusion protein shows a conformation suitable for the interferon to play functions based on the ferritin self-assembly characteristic, so that the titer of the interferon in a host is effectively improved, and the in-vivo and in-vitro half-life period of the interferon is prolonged. The invention has application prospects in preparation of drugs or reagents for preventing or treating viral diseases of grass carp and the like.

The present application relates to a method of producing a protein comprising an unnatural amino acid (UAA), the method comprising culturing a host cell, where the host cell is a eukaryotic cell, together with a nucleotide sequence encoding a first recoding tRNA or a first recoding tRNA, where the first recoding tRNA comprises an anticodon complementarily paired with a first codon, wherein the first codon is a rare codon; a nucleotide sequence encoding a first aminoacyl-tRNA synthetase or a first aminoacyl-tRNA synthetase, the first aminoacyl-tRNA synthetase being capable of loading a first recoding tRNA with a first non-natural amino acid.

The invention relates to the technical field of biological medicine, in particular to a T cell receptor targeting AFP peptide, a preparation method of the T cell receptor and a companion diagnostic kit. A T cell receptor targeting an AFP peptide is provided, the sequence of the AFP peptide is FMNKFIYEI, and the T cell receptor comprises a TCR [alpha] chain variable domain and a TCR [beta] chain variable domain. Corresponding nucleic acid molecules, vectors and host cells, and methods of making T cell receptors are provided. Further provided is a companion diagnostic kit containing the cell receptor. The invention provides a better choice for concomitant diagnosis and treatment products of cells of alpha fetoprotein (AFP).

The invention discloses a novel application of an RNF6 gene, namely, an application of screening a medicine for preventing and / or treating Zika virus infection, aiming at inhibiting expression of the RNF6 gene or knocking out the RNF6 gene. A series of functional experiments prove that the RNF6 gene promotes the replication of the Zika virus, and the purpose of reducing the replication of the Zika virus can be achieved by using an expression inhibitor of the RNF6 gene or knocking out the RNF6 gene; experimental results show that the RNF6 gene can be used as a potential target for preventing and / or treating Zika virus infection, and a new direction is provided for development of drugs for preventing and / or treating Zika virus infection targeting the RNF6 gene in the future.

The invention relates to a CHO-S cell strain capable of stably expressing H5N1 hemagglutinin protein and a construction method of the CHO-S cell strain, and belongs to the field of bioengineering.H5N1 hemagglutinin protein expression plasmids are obtained in an in-vitro synthesis and seamless cloning mode, 293T cells are transfected through the H5N1 hemagglutinin protein expression plasmids, and it is proved that the H5N1 hemagglutinin protein is expressed; the CHO-S cell strain capable of stably expressing the H5N1 hemagglutinin protein is obtained by transfecting CHO-S cells by using the H5N1 hemagglutinin protein expression plasmids and carrying out multiple rounds of cloning and screening, so that a basis is provided for obtaining H5N1 recombinant protein influenza vaccines.

The present disclosure is in the field of plant genetic engineering. Specifically, the invention relates to application of a mutant GmLn protein and an encoding gene thereof in regulation and control of soybean yield, in particular to but not limited to a method for obtaining soybeans capable of being densely planted, a method for detecting the soybeans capable of being densely planted, related primers and kits, and further relates to soybean plants capable of being densely planted and a method for improving soybean planting density. According to the method provided by the invention, the soybean target sequence is subjected to gene editing by expressing the gene editing tool in the soybean, so that the soybean which is improved in yield and can be densely planted is obtained.

The present disclosure provides compositions and methods of use thereof for targeting trans- splicing of a pre-mRNA in a cell utilizing small nuclear RNAs (snRNAs) and small nucleolar RNAs (snoRNAs), and related methods.

The invention belongs to the technical field of immunodetection, and discloses a rabbit monoclonal antibody mAb6 of an anti-mouse immune globulin G2a subtype (IgG2a) and application thereof. The rabbit monoclonal antibody mAb6 comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprises LCDR1-3, the LCDR1 comprises an amino acid sequence KSVYKNNY, the LCDR2 comprises an amino acid sequence GTN, and the LCDR3 comprises an amino acid sequence AGGYID; the heavy chain variable region comprises HCDR1-3, the HCDR1 comprises an amino acid sequence RFSLSNYR, the HCDR2 comprises an amino acid sequence IFTRGST, and the HCDR3 comprises an amino acid sequence ARGWNS. The rabbit monoclonal antibody disclosed by the invention can be combined with a mouse immune globulin G2a subtype (IgG2a) in a high-specificity manner, can be applied to immunodetection, and provides a basis for further preparation of engineering antibodies.

The invention discloses protease for improving the resistance of potatoes to late blight as well as a coding gene and application thereof, and belongs to the technical field of genetic engineering and plant disease resistance breeding. The amino acid sequence of the protease StRD21 is as shown in SEQ ID NO.2, or the protease StRD21 has a derivative sequence with the same disease-resistant function. The nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. The invention also provides a specific primer pair for cloning the gene, a potato recombinant overexpression vector containing the gene and a transgenic host cell. The StRD21 gene is introduced into potatoes and overexpressed, so that the resistance of the potatoes to late blight can be remarkably enhanced. The invention provides an effective gene resource and a biotechnological means for cultivating a new variety of disease-resistant potatoes.

The present invention relates to a method for constructing an eight-gene-edited xenogeneic organ transplantation donor pig, belonging to the field of animal biotechnology. In the wild-type porcine fetal fibroblast cell line, the GGTA1, β4GalNT2, and CMAH genes are knocked out by using the CRISPR / Cas9 gene editing technology, and the humanized genes of hCD39, hCD46, hCD55, hCD59, and hTBM are transfected. Combining with somatic cell cloning technology, GTKO / β4GalNT2KO / CMAHKO / hCD39 / hCD46 / hCD55 / hCD59 / hTBM eight-gene-edited cloned pigs are constructed. Further, through genotype, mRNA, protein expression identification and functional analysis, eight-gene-edited xenogeneic organ transplantation donor pigs are obtained. The present invention solves the technical problems of high production difficulty, low efficiency, and low survival rate of donor pigs for multi-gene-edited xenogeneic organ transplantation, and maximally solves the common problems of immune rejection reaction and complement dysregulation faced during xenogeneic organ transplantation, laying a foundation for more targeted development of donor pigs suitable for different tissue and organ xenotransplantation, and having important value for promoting the clinical transformation of xenogeneic organ transplantation.

The invention relates to a development method of an insect culture medium, which comprises the following steps: S1, constructing a double-promoter tandem vector by taking pFastBac plasmid as a skeleton; s2, establishing a baculovirus expression system; S21, transforming the double-promoter tandem vector into DH10Bac escherichia coli, so as to obtain recombinant Bacmid; s22, transfecting an Sf9 cell by using the recombinant Bacmid, so as to obtain a P0-generation baculovirus; s23, transfecting an Sf9 cell by using the P0-generation baculovirus to obtain a P1-generation baculovirus; s3, screening insect culture media: S31, providing culture media to be detected, and respectively inoculating Sf9 cells into the culture media to be detected; s32, respectively inoculating the P0-generation baculovirus or the P1-generation baculovirus into each to-be-detected culture medium for culturing; and S33, after culture is finished, detecting virus antibodies in the culture media to be detected, and screening the culture media. A comprehensive and quantitative evaluation system is established for development of the insect culture medium.

The invention belongs to the technical field of biology, and particularly relates to a mouse hepatitis virus antibody or an antigen binding fragment thereof as well as a preparation method and application thereof. The antibody or the antigen binding fragment thereof can specifically recognize and bind to the mouse hepatitis virus or the NSP3 protein thereof, and has good affinity with the mouse hepatitis virus or the NSP3 protein thereof; the kit can be used for preparing products for diagnosing, preventing and / or treating mouse hepatitis virus infection or diseases caused by the mouse hepatitis virus infection, detecting existence or level of mouse hepatitis virus or NSP3 protein in a sample, screening medicines for preventing and / or treating the mouse hepatitis virus infection or the diseases caused by the mouse hepatitis virus infection, and carrying out basic research and / or vaccine development.

The invention discloses application of MIGA2 as a therapeutic target in a medicine for preventing and treating Alzheimer's disease, and belongs to the technical field of biological medicine. The MIGA2 gene is used as a therapeutic target to be applied to development, screening or preparation of drugs for preventing and treating Alzheimer's disease. After an MIGA2 overexpression plasmid is adopted to transfect an Alzheimer's disease cell model, it is found that autophagy flow in cells can be promoted and accumulation of related toxic proteins can be reduced, it is proved that MIGA2 has the neuroprotective effect on the Alzheimer's disease, and a new way and exploration direction are provided for treatment of the Alzheimer's disease.

Disclosed herein are genetically engineered cells for AAV production. The genetically engineered cell comprises molecular systems for temporal control of expression of genes required for AAV production. Also disclosed herein are methods of using genetically engineered cells for AAV production.

The invention relates to a monoclonal antibody with anticoagulant activity and application thereof, the antibody or fragment comprises a light chain variable region and a heavy chain variable region, and the amino acid sequence of the light chain variable region of the antibody or fragment is as shown in SEQ ID NO: 1; the amino acid sequence of the variable region of the heavy chain is as shown in SEQ ID NO: 3. The monoclonal antibody disclosed by the invention has the effect of inhibiting the activity of a co-coagulation pathway in a coagulation cascade reaction, and can be applied to prevention and treatment of thrombotic diseases.

Polynucleotides, vectors, systems of vectors or polynucleotides, cells, and methods for expressing AAV Rep and Cap proteins are provided. In certain aspects, these polynucleotides, cells, and methods may produce higher levels of one or both of Rep and Cap transcripts. In some aspects, these polynucleotides, cells, and methods may produce higher levels of one or both of Rep and Cap proteins. In various aspects, these polynucleotides, vectors, systems of vectors or polynucleotides, cells, and methods use one or more heterologous polyA sequences with one or both of the Rep and Cap coding sequences. In numerous aspects, these polynucleotides, vectors, systems of vectors or polynucleotides, cells, and methods may use one or more enhancers to increase expression of Rep proteins, where such enhancers can be transcriptional enhancers, translational enhancers, or combinations thereof. In certain aspects, these polynucleotides, vectors, systems of vectors or polynucleotides, cells, and methods may be used to produce recombinant AAV (rAAV).

The invention belongs to the technical field of immunodetection, and discloses an anti-T3 antibody mAb32 or an antigen binding fragment, the anti-T3 antibody mAb32 or the antigen binding fragment comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprises LCDR1, LCDR2 and LCDR3, the amino acid sequence of the LCDR1 is shown as SEQ ID NO.1, the amino acid sequence of the LCDR2 is IAS, and the amino acid sequence of the LCDR3 is shown as SEQ ID NO.3; the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the amino acid sequences of the HCDR1, the HCDR2 and the HCDR3 are respectively shown as SEQ ID NO. 4-6. When the anti-T3 antibody mAb32 is used for detecting a T3 standard antigen, the detection sensitivity is lower than 0.2 ng / ml, and when a clinical sample is detected through a magnetochemiluminescence method, the correlation with R clinical comparison is good within the sample range of 1-80 pmol / L.