272 results about "Eukaryotic cell" patented technology

The invention discloses a small CRISPR (clustered regularly interspaced short palindromic repeats)-Cas gene editing system and application thereof. According to the invention, based on microbial genome and metagenome data, a class of CRISPR-Cas family protein is mined through a biological information method, and is named as Cas12r. A CRISPR-Cas12r editing tool constructed on the basis of the gene can realize gene editing in prokaryotic or eukaryotic cells. The CRISPR-Cas12r gene editing system obtained by the invention has the characteristics of miniaturization and various PAM types.

The invention discloses a recombinant protein for detecting a para-tumor Yo antibody through a CBA method and application, and belongs to the technical field of biomedical engineering.The amino acid sequence of the recombinant protein sequentially comprises a secretory signal peptide, a CDR2 protein partial sequence, a CDR2L protein partial sequence, a transmembrane region and a fluorescent label, and the secretory signal peptide is human albumin signal peptide ALB; the transmembrane region is a CD8a hinge; and the fluorescent label is mCherry. A novel recombinant protein which can be stably over-expressed on a cell membrane of an eukaryotic cell is constructed by intercepting specific partial sequences of CDR2 protein and CDR2L protein and redesigning and fusing a fluorescent label by using a secretory signal peptide, a transmembrane sequence and a connecting peptide, and the recombinant protein retains respective core antigen regions of the CDR2 protein and the CDR2L protein, so that the specific partial sequences of the CDR2 protein and the CDR2L protein can be stably over-expressed on the cell membrane of the eukaryotic cell. The kit can effectively overcome the defects in the aspects of sensitivity and specificity, and when a CBA method is adopted for detection, the detection rate of the para-tumor Yo antibody can be remarkably increased, and the false positive rate is reduced, so that the requirements of clinical detection are better met.

The invention discloses a human-derived anti-respiratory syncytial virus neutralizing antibody RS135 and application of the human-derived anti-respiratory syncytial virus neutralizing antibody RS135. The human neutralizing antibody specifically aiming at the F protein of the respiratory syncytial virus is successfully obtained by applying a phage antibody library technology; the obtained human neutralizing anti-respiratory syncytial virus F protein gene engineering antibody variable region gene, Fab antibody gene and full antibody gene under the characteristics of each antibody gene are utilized to obtain the human neutralizing anti-respiratory syncytial virus F protein gene engineering antibody. The antibody can be expressed and produced in prokaryotic cells, yeast cells, eukaryotic cells and any recombination system or any other reconstructed gene containing the antibody gene on the basis of the antibody, and an antibody product capable of neutralizing respiratory syncytial virus infection is obtained. And a specific antibody drug for clinically preventing and treating respiratory syncytial virus infection is prepared.

The application provides an in-vitro cell-free protein synthesis system and kit containing exogenous magnesium ions and application thereof, including a D2P system (DNA-to-Protein system) and an mR2P system (mRNA-to-Protein system), and belongs to the technical field of protein synthesis. The in-vitro cell-free protein synthesis system containing exogenous magnesium ions adopts magnesium aspartate as a novel magnesium ion source, and especially in a eukaryotic cell-free system, compared with a traditional magnesium ion source, can significantly improve the protein synthesis efficiency and protein expression amount. A more efficient and higher-throughput in-vitro protein synthesis kit and a synthesis method of exogenous proteins are also provided, and the kit has the advantages of simplicity, convenience and low cost.

The invention relates to a vaccine for preventing and treating sheep echinococcosis infection and a preparation method thereof.The preparation method comprises the steps that echinococcosis Eg95 full-length gene antigen epitopes are screened through bioinformatics software, screened fragments with high antigenicity are optimized and then subjected to double-copy connection through a flexible linker, His tag sequences are added to the two ends of a tandem sequence, the sequence is as shown in SEQ ID NO.2, and the vaccine for preventing and treating sheep echinococcosis infection is obtained; and the pEE12.4-double Eg95 plasmid is connected with a CHO eukaryotic cell expression vector, so that the pEE12.4-double Eg95 plasmid is obtained. The plasmid is integrated into a CHO eukaryotic cell expression system, the expression quantity of the purified Eg95 protein reaches 10g / L, and a stable and high-yield veterinary vaccine is provided for large-scale production.

The present invention relates to the technical field of biomedicine. Specifically provided are a respiratory syncytial virus (RSV) Pre-F recombinant protein nanoparticle vaccine and a preparation method therefor. By performing amino acid mutation on an RSV Pre-F protein, and performing fusion expression on the mutated Pre-F protein and ferritin particles in eukaryotic cells, PreF-Ferritin fusion protein nanoparticles, in which eight Pre-F protein trimers are densely displayed on the surface of each ferritin particle, are obtained. By stabilizing and exposing antigen epitopes required to be displayed and disrupting or masking unrequired antigen epitopes, the PreF-Ferritin fusion protein nanoparticles effectively improve the immunogenicity and the production stability of the antigen. Experiments show that: when the PreF-Ferritin fusion protein is injected into mice, high-titer protective sera can be acquired, and the mice sera can yield relatively high neutralizing titers against RSV.

The present invention relates to nucleic acids coding for a protein said protein comprising a truncated glial fibrillary acidic protein (GFAP), proteins comprising a truncated GFAP, expression constructs comprising a nucleic acid coding for a protein which comprises a truncated GFAP, vectors comprising the nucleic acids or expression constructs disclosed herein and eukaryotic cells comprising a nucleic acid, proteins, expression construct or vector disclosed herein. The present invention furthermore relates to compositions comprising the herein disclosed proteins and kits comprising the herein disclosed proteins and / or compositions. Furthermore, the present invention relates to the use of the herein disclosed proteins, compositions and / or kits for calibrating a measurement system for the in vitro quantification of GFAP or a variant thereof in a sample.

Described herein are compositions and methods for modifying eukaryotic cells, for example, to express a transgene of interest and / or to produce an expanded population of cells ex vivo. Using the compositions and methods of the disclosure, a population of eukaryotic cells, such as a population of pluripotent cells (e.g., CD34+ hematopoietic stem or progenitor cells) may be transduced to express a gene of interest by contacting the cells with a viral vector, such as a lentiviral vector, and a diblock copolymer, such as a diblock copolymer composed of a hydrophilic region and a hydrophobic region. For example, the diblock copolymer may be composed of polyoxyethylene (PEO) subunits and polyoxypropylene (PRO) subunits. Additionally, the compositions and methods described herein can be used to promote the proliferation or survival of a population of pluripotent cells (e.g., CD34+ hematopoietic stem or progenitor cells) ex vivo, for example, by contacting the cells with a diblock copolymer.

Methods and compositions are provided for the improvement of double-strand-break-inducing agent activity in eukaryotic cells, through the usage of one or more morphogenic factors or developmental genes. The morphogenic factor may be provided to the same cell or to a different cell than that comprising or receiving the double-strand-break-inducing agent. The morphogenic factor may be provided to a cell as a polynucleotide composition on a recombinant vector, and may be placed on the vector outside of a T-DNA border. The morphogenic factor may be provided via an upregulation of an endogenous gene. The morphogenic factor, or the double-strand-break-inducing agent, may further comprise a cell penetrating peptide. The morphogenic factor may be co-introduced with a vector comprising RepA.

This application relates to a method for producing a protein containing an unnatural amino acid (UAA), the method comprising culturing a host cell, wherein the host cell is a eukaryotic cell, together with: a nucleotide sequence encoding a first recoding tRNA or a first recoding tRNA, wherein the first recoding tRNA comprises an anticodon complementary to a first codon, and wherein the first codon is a rare codon; and a nucleotide sequence encoding a first aminoacyl-tRNA synthetase or a first aminoacyl-tRNA synthetase, wherein the first aminoacyl-tRNA synthetase is capable of charging the first recoding tRNA with the first unnatural amino acid.

Described herein are methods, compositions, kits, and synthetic peptide shuttles involving transduction of protein and / or non-protein cargoes. The methods generally comprise contacting a target eukaryotic cell with a non-protein cargo and a concentration of a synthetic peptide shuttle sufficient to increase the transduction efficiency of the non-protein cargo as compared to the absence of the synthetic peptide shuttle. In embodiments, the non-protein cargo can be a drug, such as a small molecule drug, for treating a disease. In other embodiments, novel synthetic peptide shuttles active in transducing protein and / or non-protein cargoes are described, as well as the use of propidium iodide or other membrane-impermeable fluorescent DNA intercalators as surrogate cargoes for selecting multifunctional synthetic peptide shuttles active in transducing both protein and non-protein cargoes.

The present disclosure relates to modulation of protein expression.

The present invention relates to a recombinant antisense oligonucleotide that targets CD33 mRNA and to a method of preparing a substantially pure population of edited eukaryotic cells comprising the steps of i) editing a population of eukaryotic cells by the use of base editors, (ii) treating the same population with antisense oligonucleotides according to the invention to transiently downregulate CD33 and iii) enriching the population of edited eukaryotic cells that is negative for CD33.

The present invention relates to a nucleic acid molecule encoding (I) a polypeptide having the activity of an endonuclease, which is (a) a nucleic acid molecule encoding a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1; (b) a nucleic acid molecule comprising or consisting of the nucleotide sequence of SEQ ID NO: 2; (c) a nucleic acid molecule encoding an endonuclease, the amino acid sequence of which is at least 70% identical to the amino acid sequence of SEQ ID NO: 1; (d) a nucleic acid molecule comprising or consisting of a nucleotide sequence which is at least 50% identical to the nucleotide sequence of SEQ ID NO: 2; (e) a nucleic acid molecule which is degenerate with respect to the nucleic acid molecule of (d); or (f) a nucleic acid molecule corresponding to the nucleic acid molecule of any one of (a) to (e) wherein T is replaced by U; (II) a fragment of the polypeptide of (I) having the activity of an endonuclease. Also, the present invention relates to a vector comprising the nucleic acid molecule and a protein encoded by said nucleic acid molecule. Further, the invention relates to a method of modifying the genome of a eukaryotic cell and a method of producing a non-human vertebrate or mammal.

The invention provides a novel base editor and application thereof. Specifically, the invention provides a novel double-stranded DNA deaminase, and a fusion protein (novel base editor), a base editing system, a base editing method and the like based on the novel double-stranded DNA deaminase, so as to improve the DNA editing efficiency of eukaryotic cell genes (especially mitochondrial genes), and especially improve the editing efficiency of non-TC DNA fragments such as' AC ',' GC 'and the like. According to the method, the types of the DNA fragments which can be effectively edited are expanded, and a wider prospect is provided for clinical application of DNA editing.

The invention relates to a composition for regulating autophagy of eukaryotic cells and improving mitochondrial dysfunction as well as a preparation method and application of the composition. Comprising the following raw materials in parts by weight: 10-30 parts of broccoli powder, 10-30 parts of wheat germ microcapsule powder, 20-40 parts of raspberry extract, 20-40 parts of banana powder, 2-6 parts of leucine and 0.1-1 part of pyrroloquinoline quinone disodium salt. The invention further provides a preparation method of the broccoli powder, the wheat germ micro-capsule powder, the raspberry extract and the banana powder, and application of the composition of the broccoli powder, the wheat germ micro-capsule powder, the raspberry extract and the banana powder. The composition integrates the effects of the six components, is reasonable in compatibility, safe and effective, and various components activate mitochondria by regulating protein and depending on a PINK1 / Parkin dependent pathway and a PINK1 / Parkin non-dependent pathway; meanwhile, by activating a PGC-1alpha signal channel, mitochondrial DNA replication and new mitochondrial generation are promoted; the mitochondrial membrane potential is stabilized, and the ATP synthesis efficiency is improved; mitochondrial autophagy is synergistically activated, mitochondrial dysfunction is improved, and aging is relieved.

The invention discloses an editing system for repairing amyotrophic lateral sclerosis pathogenic mutant genes, and relates to an IS621-bridge-RNA gene editing system capable of being applied to eukaryotic cells, an AAV9 virus vector mediated IS621-bridge-RNA gene editing system and application of the AAV9 virus vector mediated IS621-bridge-RNA gene editing system, and the IS621-bridge-RNA gene editing system, a recombinant expression vector and a gene therapy mode are included.

The application discloses a CRP camel-derived single-domain antibody and a preparation method and application thereof, relates to the field of biological medicines, and is characterized in that: white blood cells of a llama immunized with a CRP antigen are enriched, and then single-cell screening and sequencing are performed. Through sequencing structure analysis, a camel-derived CRP antibody recombinant plasmid is constructed, a related antibody is expressed in a eukaryotic cell through cloning transformation, a single antibody with human CRP resistance is screened, a VHH domain of the single antibody is cloned and expressed, and a single antibody with CRP-VHH antibody activity is obtained through pairing screening. The antibody can be applied to an ELISA kit, a chemiluminescence kit and a latex kit, has the same functions as existing kits, and still has the use in the preparation of an anti-tumor drug.

The present invention relates to nucleic acids coding for a protein said protein comprising a glial fibrillary acidic protein (GFAP) and a eukaryotic secretion tag, proteins comprising a GFAP and a eukaryotic secretion tag, expression constructs comprising a nucleic acid coding for a protein which comprises a GFAP and a eukaryotic secretion tag, vectors comprising the nucleic acids or expression constructs disclosed herein and eukaryotic cells comprising a nucleic acid, expression construct or vector disclosed herein. The present invention furthermore relates to methods for the production of the herein disclosed proteins. The present invention also relates to compositions comprising the herein disclosed proteins and kits comprising the herein disclosed proteins and / or compositions. Furthermore, the present invention relates to the use of the herein disclosed proteins, compositions and / or kits for calibrating a measurement system for the in vitro quantification of GFAP or a variant thereof in a sample.

The present disclosure concerns polynucleotides and amino acids of Acidaminococcussp. Casl2a (Cpfl) and methods for their use for genome editing in eukaryotic cells. 20 24 20 14 49 05 M ar 2 02 4 A B S T R A C T 2 0 2 4 2 0 1 4 4 9 0 5 M a r 2 0 2 4

The application provides a kind of mitochondrial function optimization additive for cell culture, preparation method and application, belongs to cell culture technical field.The additive includes core active ingredient, auxiliary component and carrier, and the core active ingredient includes D-Arg-Cha-Lys-Phe-NH2 Polypeptide, ubiquinone-10 phosphate, reduced glutathione and alpha-lipoic acid.The application forms a multi-target core active system through the synergy of the four, realizes the triple synergistic protection of stabilizing mitochondrial cardiolipin, maintaining mitochondrial morphology and membrane potential, and reducing mitochondrial ROS, which can significantly improve cell culture quality.When applied, the mitochondrial cardiolipin content can be increased by more than 50%, the mitochondrial abnormality ratio can be reduced to less than 12%, the membrane potential positive cell ratio can be increased to more than 86%, the mitochondrial ROS level can be reduced by more than 40%, and the cell proliferation rate can be increased by more than 25%, which is suitable for in vitro culture of various eukaryotic cells, and the preparation process is simple and has no cytotoxicity.

Provided herein is a eukaryotic cell expressing an exogenously introduced inosine 5'-monophosphate dehydrogenase (IMPDH) that is resistant to a purine biosynthesis inhibitor, uses thereof for producing therapeutic agents and treating a disease or disorder, as well as pharmaceutical compositions and methods of making thereof.

A fusion protein for an auxin degradation determining subsystem for controlling the degradation of a target protein derived from a non-plant eukaryotic cell, the fusion protein being a fusion protein of a degradation tag and a protein having an affinity for the target protein, the dissociation tag comprises at least a portion of the Aux / IAA family protein and has an affinity for the TIR1 family protein-auxin or auxin analog complex.

The construct for site-specific insertion of DNA fragments and transgenes into the genome comprises a recombinant enzyme. This enzyme contains a modified HIV-1 integrase (HIV IN) tetramer with multiple mutations favoring a heterodimer structure in which the first and second outer monomers are bound to the first and second inner monomers, respectively. A transcription activator-like effector (TALE) is bound to the N-terminal region of the outer monomer via a polypeptide linker. The two TALEs confer sequence specificity to the enzyme and prevent non-specific interactions with the target DNA structure. The modified HIV IN monomer ensures that the TALE binds to the outermost monomer and that the TALE is positioned relative to the catalytic region of HIV IN. A polynucleotide is bound to the inner monomer of HIV IN. When the TALE is transported to the nucleus, either alone or within a lentiviral capsid, it interacts with the DNA to localize HIV IN, which then incorporates the polynucleotide.

CRISPR-Cas systems have been engineered for various purposes, such as genomic DNA cleavage, base editing, epigenome editing, and genomic imaging. Although significant developments have been made, there still remains a need for new and useful CRISPR-Cas systems as powerful precise genome targeting tools. The invention disclosed herein comprises CRISPR-Cas based compositions and methods for high integration efficiency and expression efficiency of transgenes together with high post-transfection cell viability in eukaryotic cells.

The present invention provides polynucleotide vectors for high expression of heterologous genes. Some vectors further comprise novel elements that further improve expression. The gene transfer systems can be used in methods, for example, gene expression, bioprocessing, gene therapy, insertional mutagenesis, or gene discovery.

The application discloses a gene editing system CRISPR-Cas12p and application thereof. Based on microbial genomes and metagenomic data, the CRISPR-Cas12p protein of the CRISPR-Cas protein family is obtained by preliminary screening by using a Prodigal gene prediction tool, a Pfam database and HMMER software, and a gene editing system CRISPR-Cas12p is constructed. PAM preference and interference function identification show that the editing system has a PAM preference of 5'-TTC-3', can effectively realize targeted cutting by using long transcripts and double RNA hybrid chain transcripts respectively, and can realize editing of a target gene in prokaryotic and eukaryotic cells. The CRISPR-Cas12p gene editing system obtained by the application has a small protein component, is beneficial to delivery, can realize gene editing in prokaryotic and eukaryotic cells, and has a wide application prospect.

The present disclosure provides compositions and methods of inactivating a virus in a product feedstream in a manufacturing process of a therapeutic agent using an antifoam reagent. In some aspects, the therapeutic agent comprises a therapeutic protein, a viral particle, a nucleic acid molecule, or any combination thereof. Other aspects of the present disclosure are directed to methods of inactivating a virus in a eukaryotic cell culture of a product feedstream, comprising contacting the cell culture with an antifoam reagent. In some aspects, the antifoam reagent comprises an emulsifier, e.g., a nonionic emulsifier, and optionally a silicone polymer.

The preparation of a library of bispecific binders expressed in eukaryotic cells described herein is a method for producing a cloned library of eukaryotic cells containing DNA encoding a diverse library of bispecific binders, the present invention relates to bispecific conjugates, in particular bispecific conjugates comprising a first binding domain coupled to a first Fc domain and a second binding domain coupled to a second Fc domain.

Provided are a truncated chromatin opening element and a combination thereof, an expression system comprising the chromatin opening element or the combination thereof and applied to eukaryotic cells, and use thereof. The chromatin opening elements are truncated sequences of two chromatin opening elements from different sources, respectively, and can be used alone or in combination. The chromatin opening element significantly shortens the effective sequence, thereby facilitating the integration of a vector comprising the element or the combination thereof with other elements, such as integration with ITR, achieving semi-site-specific integration in the presence of the PiggyBac transposase, and then using a bulkpool rapid screening process to shorten the protein production and cell strain construction cycle by about 4-5 weeks while maintaining comparable yields.