383 results about "Eukaryotic cell" patented technology

The invention relates to retrieving or modifying target nucleic acids, such as host cell chromosomal DNA, by homologous recombination with vectors that have been cut to provide recombinogenic nucleic acid strands in situ. The methods described herein can be used to modify a target nucleic acid in vitro or in any prokaryotic or eukaryotic cell using homologous recombination.

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 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.

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 methods for high integration and expression efficiency of transgenes together with high post-transfection cell viability in eukaryotic cells.

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.

The invention provides functional identification and application of a novel cell virulence factor. Based on the research on the novel virulence factor of the cell, the invention also provides a polypeptide delivery system and application thereof. Wherein the polypeptide delivery system comprises a protein complex and a signal peptide, the signal peptide comprises at least 20 amino acids at the N-terminal of PagT1 protein or homologous protein thereof, and the signal peptide is used for guiding the polypeptide to be loaded into the protein complex and guiding the polypeptide to be positioned to the nucleus of eukaryotic cells. The research of the invention provides an important basis for constructing a safe and efficient novel tumor treatment system, and provides an innovative solution for treating serious diseases such as cancers.

Use of impedance and / or dielectric spectroscopy for analysis of a eukaryotic cell expressing an endogenous or recombinant peptide or polypeptide. A sensing apparatus 30 includes a main channel 34 thro

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.

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.

The invention relates to a method to determine a homology directed repair (HDR) event within a eukaryotic cell, wherein the cell expresses a first isoform of a surface protein, which is different from a second isoform of said surface protein with regard to an amino acid marker. The method comprises the steps of inducing a DNA double strand break, providing a HDR template DNA construct comprising the amino acid marker corresponding to the second isoform of the surface protein and subsequently determining the expression of the first or second isoform of said surface protein on said cell, wherein expression of the second isoform indicates a successful HDR event. The invention also relates to a method for editing a genomic location of interest within a eukaryotic cell, and to a method of selectively depleting or enriching an edited cell in a composition of non-edited and edited cells.

The present inventions provide eukaryotic cells, such as mammalian cells, that comprise adeno-associated virus (AAV) polynucleotides, including AAV capsid proteins (Cap), and are capable of expressing the polypeptides encoded by the AAV polynucleotides, and thereby are capable of producing AAV, including recombinant AAV. The eukaryotic cells also may comprise adenovirus (Ad) polynucleotides. The present inventions also provide methods of expressing AAV polynucleotides, as well as Ad polynucleotides, in eukaryotic cells, such as CHO cells, HEK 293 and BHK cells. The present inventions further provides other products and methods described herein.

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.

The invention provides a truncated insulator and an expression system comprising the insulator and applied to eukaryotic cells, the truncated insulator breaks the setting of an original core area, shortens the effective sequence of the insulator, and is more beneficial to the integration of the insulator and other elements of a carrier. The insulator can obviously improve the yield of protein in an expression system with transposon (such as PiggyBac) and obtain a recombinant cell strain with good hereditary stability.

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.

Engineered Cas9 systems that utilize alternate protospacer adjacent motifs for target DNA binding, nucleic acids encoding the engineered Cas9 systems, and methods of using the engineered Cas9 systems for modifying target chromosomal sequences in eukaryotic cells.

A method for modifying a gene encoding or processed to a non-coding RNA molecule having no RNA silencing activity in a eukaryotic cell is disclosed, with the proviso that the eukaryotic cell is not a plant cell. The method comprises introducing a DNA editor into the eukaryotic cell, the DNA editor conferring a silence specificity of the non-coding RNA molecule for a target RNA of interest. Also disclosed is a method of modifying a gene for encoding or being processed into an RNA silencing molecule for a target RNA in a eukaryotic cell. Also disclosed are methods of preventing and treating various diseases, methods of inducing apoptosis, and methods of producing an eukaryotic non-human organism.

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 targeted genome editing in eukaryotic cells or organisms. More particularly, the present invention relates to a composition for cleaving a target DNA in eukaryotic cells or organisms comprising a guide RNA specific for the target DNA and Cas9 protein, and uses thereof.

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 invention discloses a CRISPRa tool for activating a target gene and application of the CRISPRa tool. The invention provides a protein. The protein sequentially comprises the following elements from an N terminal to a C terminal: a dCas9 segment and an activation domain segment, n NHRs are arranged in the active domain section; n is a natural number; the NHR is an NHR activating element from a nematode. The NHR has a nonaamino acid trans-activation domain (9aa TAD), consists of only 16 amino acid residues, and shows high transcriptional activity. The invention provides a CRISPRa tool with stronger action effect and more concise action element, can be used for gene transcription regulation (particularly gene transcription activation), can be used in the scientific research field and the clinical medicine field, and has great value.