167 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 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 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 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 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.

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 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.

The invention relates to peptide-based non-protein load delivery. Methods, compositions, kits and synthetic peptide shuttle agents related to transduction of protein and / or non-protein loads are described. The methods generally include contacting a target eukaryotic cell with a non-protein load and a synthetic peptide shuttle at a concentration sufficient to increase the transduction efficiency of the non-protein load compared to the absence of the synthetic peptide shuttle. The non-protein load may be a drug for the treatment of disease, such as a small molecule drug. The invention also describes novel synthetic peptide shuttles having transduction activity on protein and / or non-protein loads, as well as the use of propidium iodide or other membrane impermeable fluorescent DNA intercalators as alternative loads for selecting multifunctional synthetic peptide shuttles having transduction activity on both protein and non-protein loads.

The present invention is directed to a method for identifying i) metastatic potential of a cancer cell sample from a non-metastatic cancer or ii) potential for malignant alterations in a healthy cell, the method comprising the steps of: a) culturing a cancer cell sample from a non- metastatic cancer or a healthy cell sample in a composition comprising cancer-associated fibroblasts, CAFs, preferably in two-dimensional or three-dimensional cell culturing conditions; and b) monitoring morphological alterations in the sample cells during and / or after the culturing step a), and / or determining the expression level of biomarkers relating to metastasis in the sample cells during and / or after the culturing step a); wherein the sample cells which show morphological alterations relating to metastasis and / or changes in the expression levels of said biomarkers in comparison to the levels of said biomarkers measured from corresponding samples cultured in a composition comprising normal fibroblasts, NFs, are identified as having metastatic potential or potential for malignant alterations. The present invention is also directed to a system for culturing eukaryotic cells comprising a solid surface comprising at least one partition suitable for culturing eukaryotic cells, wherein said at least one partition contains a combined cell culture comprising i) cancer cells from a non- metastatic cancer or healthy cells and ii) cancer-associated fibroblasts, CAFs, wherein said partition is a well or chamber.

This provides an inhibitor of protein fucosylation. [Solution] A method for inhibiting the fucosylation of a protein, or a fragment or derivative thereof, comprising contacting a eukaryotic cell or mammal with a specific carbocyclic compound or a salt thereof.

The application provides a kind of mitochondrial function optimization additive for cell culture, preparation method and application, belong 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.

A method of altering a eukaryotic cell is provided including transfecting the eukaryotic cell with a nucleic acid encoding RNA complementary to genomic DNA of the eukaryotic cell, transfecting the eukaryotic cell with a nucleic acid encoding an enzyme that interacts with the RNA and cleaves the genomic DNA in a site specific manner, wherein the cell expresses the RNA and the enzyme, the RNA binds to complementary genomic DNA and the enzyme cleaves the genomic DNA in a site specific manner.

An auxin-inducible degron system kit that controls degradation of a target protein in a non-plant-derived eukaryotic cell, the kit containing a first nucleic acid that encodes a mutant TIR1 family protein having a mutation at an auxin-binding site, an auxin analog that has an affinity to the mutant TIR1 family protein and a second nucleic acid that encodes a degradation tag containing at least a part of an Aux / IAA family protein and having an affinity to a complex of the mutant TIR1 family protein and the auxin analog.

The present disclosure relates to targeted genome editing in eukaryotic cells or organisms. More particularly, the present disclosure provides for compositions and methods that may induce modifications in target endogenous nucleic acid sequences in nucleuses of eukaryotic cells. For example, disclosed herein is a method of producing an engineered eukaryotic cell. In some embodiments, the method may comprise preparing a Cas9 protein, preparing a sgRNA, and preparing a cell-free buffer. The method may further comprise disposing the Cas9 protein and sgRNA in the cell-free buffer to provide for a transfection mixture and transfecting the transfection mixture into a eukaryotic cell, wherein a Cas9 / sgRNA complex formed by the Cas9 protein and the sgRNA induces a modification of a target endogenous DNA sequence in the nucleus of the eukaryotic cell to provide for an engineered eukaryotic cell.

Method of increasing protein content in a eukaryotic cell comprising an NF-YC4 gene comprising modifying the transcriptional repressor binding site; method of producing a plant with increased protein content comprising crossing and selecting for increased protein content; method of increasing resistance to a pathogen or a pest in a plant cell or plant comprising an NF-YC4 gene comprising modifying the transcriptional repressor binding site, alone or in further combination with expressing QQS in the plant cell or plant; method for producing a plant with increased resistance to a pathogen or a pest comprising crossing and selecting for increased resistance to the pathogen or the pest; a cell, collection of cells, tissue, organ, or organism, such as a plant, in which the NF-YC4 gene comprises a promoter comprising a transcriptional repressor binding site that has been modified so that the transcriptional repressor cannot prevent transcription of the NF-YC4; plants and hybrids thereof; and seeds.

One objective is to provide a means for intracellular delivery of non-protein cargo. [Solution] Methods, compositions, kits, and synthetic peptide shuttles relating to the transduction of protein cargoes and / or non-protein cargoes are described herein. The methods generally include contacting target eukaryotic cells with a synthetic peptide shuttle at a concentration sufficient to increase the transduction efficiency of the non-protein cargo compared to the absence of the non-protein cargo and the synthetic peptide shuttle. In embodiments, the non-protein cargo may be a drug, such as a small molecule drug for treating a disease. In other embodiments, novel synthetic peptide shuttles having transduction activity for protein cargoes and / or non-protein cargoes are described, and further, the use of propidium iodide or other membrane-impermeable fluorescent DNA insertion agents as surrogate cargoes for selecting a versatile synthetic peptide shuttle having transduction activity for both protein and non-protein cargoes is described.

Provided herein are systems, methods, and compositions for the modification of target DNA sequences. More particularly, systems, methods, and compositions for cleaving a target DNA in eukaryotic cells with a guide RNA capable of hybridizing with a target sequence and an RNA-guided DNA nuclease are provided. Also provided are vectors and vector systems which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods for identifying and validating novel CRISPR systems.

This document relates to methods and materials for genome engineering in eukaryotic cells, and particularly to methods for increasing genome engineering (i.e. transformation or genome editing) efficiency via delivery of one or more RKD2 and RKD4 genes, with genome engineering components.

Compositions and methods are provided for modifying eukaryotic cells, e.g., to express a transgene of interest and / or to generate a population of expanded cells ex vivo.SOLUTION: A method of transducing a population of eukaryotic cells, such as a population of pluripotent cells, to express a gene of interest by contacting the cells with a viral vector, e.g., a lentiviral vector, and a diblock copolymer, e.g., a diblock copolymer comprised of a hydrophilic region and a hydrophobic region. For example, the diblock copolymer can be composed of polyoxyethylene (PEO) subunits and polyoxypropylene (PRO) subunits. In addition, the compositions and methods of the invention can be used to promote the ex vivo expansion or survival of a population of pluripotent cells (e.g., CD34 + hematopoietic stem or progenitor cells), for example, by contacting the cells with a diblock copolymer.SELECTED DRAWING: None

The invention relates to methods for the flexible regulation of cellular growth of eukaryotic and prokaryotic cells. In particular embodiments, regulation of eucaryotic and microbial cells growth, occurs by the control of their interaction with the environmental factors, nutrient media, media additives, supplements.

Provided are an RSV vaccine as well as a preparation method therefor and the use thereof. A Pre-F related sequence of an RSV and ferritin nanoparticles are subjected to mutation design, and a Pre-F mutant protein and a ferritin mutant are fused and expressed in eukaryotic cells, so as to obtain ferritin-PreF fusion protein nanoparticles having multiple Pre-F displayed on the surfaces in a centralized manner, and the amino acid sequence of the ferritin-PreF fusion protein nanoparticles is any one of SEQ ID No. 20-27.