527 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 provides a construction method and application of a high-efficiency expression system based on a Seali forest virus replicon. The method comprises the following steps: constructing a saRNA system for expressing a reporter gene; constructing to obtain replicase providing plasmids in the taRNA system; constructing to obtain an antigen providing plasmid in the taRNA system; the method comprises the following steps: respectively linearizing pSFV-CMV-T7-X, pSFV-CMV-T7-TR-X and pSFV-CMV-T7-replicase, and then carrying out in vitro transcription to prepare mRNA (Messenger Ribonucleic Acid); transfecting the prepared mRNA into a host eukaryotic cell for expression according to a certain proportion to obtain an antigen protein; and detecting the antigen protein to evaluate the expression efficiency of the saRNA and taRNA system based on the SFV replicon. According to the self-amplification DNA and RNA vaccine based on the virus replicon, the development period can be shortened, and the vaccine antigen can be expressed at a high level.

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 present invention relates generally to the field of site-specific DNA recombination mediated by lambda integrase, and more specifically, to a method for producing eukaryotic cells and cell lines comprising a genomic landing pad for lambda integrase-mediated recombination, and methods of using the eukaryotic cells themselves and subsequently their use in lambda integrase mediated recombination and as bioreactors for cell therapy.

The invention relates to antisense oligonucleotides that are capable of bringing about specific editing of a target nucleotide (adenosine) in a target RNA sequence in a eukaryotic cell, wherein said oligonucleotide does not, in itself, form an intramolecular hairpin or stem-loop structure, and wherein said oligonucleotide comprises a non-complementary nucleotide in a position opposite to the nucleotide to be edited in the target RNA sequence.

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.

Compositions of bacterial tyrosyl-tRNA synthetase mutants that enable site-specific incorporation of numerous unnatural amino acids into proteins expressed in eukaryotic cells or in engineered ATM E. coli and methods of use are described. Compositions of novel unnatural amino acids that can be incorporated using these mutant bacterial tyrosyl-tRNA synthetase are also described.

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.

Compositions and methods for using programmable DNA binding proteins to increase the efficiency and / or specificity of targeted genome modification or to facilitate the detection of specific genomic loci in eukaryotic cells.

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 present invention relates to an antisense long non-encoding RNA (aslncRNA) that may form a partial double stranded (ds) complex with a corresponding cellular mRNA in eukaryotic cells and to a biological complex comprising the antisense lncRNA and a target mRNA that may together form a partial double stranded complex in vitro. The dsRNA complex is preferentially exported from the nucleus and translated in the cytoplasm and thereby gene expression is boosted. In addition to boosting aslnc-RNAs inhibitiory aslnc RNAs are disclosed. An expression vector comprising a nucleic acid sequence of the aslncRNA is also disclosed. The present invention can be used to increase or decrease export of a target mRNA from the nucleus and thus, to increase or decrease gene expression without interfering with genomic DNA or with mRNA levels. Methods to increase or decrease the export and / or expression of a target mRNA, cell transfections using plasmids expressing the antisense lncRNA or using conjugates comprising the aslncRNA, and a pharmaceutical composition comprising the as-lncRNA are also disclosed. The aslncRNA, the complex, the expression vector, and the conjugate are useful for enhancing the production of recombinant proteins that are used in biotechnological applications and for medical purposes. The aslncRNA is also useful for increasing antigen presentation upon mRNA vaccination.

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.

The application relates to the field of oncology, cancer immunotherapy, molecular biology and recombinant nucleic acid technology. In particular, the invention relates to genetically-modified eukaryotic cells comprising modulated TGF Beta signaling. The application further relates to the use of such genetically-modified eukaryotic cells for treating a disease, including cancer, in a subject.

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.

In vitro or ex vivo methods for screening a quinoline derivative, or anyone of its pharmaceutically acceptable salt, presumed effective in treating and / or preventing an inflammatory disease, the method including: providing an eukaryotic cell, bringing into contact said cell with a quinoline derivative, measuring an expression of miR-124 in the cell, and selecting the candidate presumed effective in treating and / or preventing an inflammatory disease when the level of expression of miR-124 measured is increased relatively to a reference value.

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.

Ionizable lipids are provided that are useful for delivering macromolecules, such as nucleic acids, into eukaryotic cells. The lipids can be used alone, in combination with other lipids and / or in combination with other transfection enhancing reagents to prepare transfection complexes.

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.