55 results about "Replication cycle" patented technology

The invention belongs to the field of biological medicines, and discloses an application of a VPS35 inhibitor in preparation of Zika virus infection resistant drugs, the VPS35 inhibitor comprises at least one of the following two substances: I: a substance for inhibiting VPS35 gene expression in host cells; and II: a substance for inhibiting the retrograde transport function of the VPS35 protein in the host cell. The invention finds that the VPS35 protein is an important host factor of the replication period of the Zika virus for the first time, and a Zika virus infection model system verifies that the replication of the Zika virus in the host cell can be inhibited by inhibiting the expression of the VPS35 gene in the host cell or inhibiting the retrograde transport function of the VPS35 protein. Compared with traditional drugs targeting Zika virus protein, the VPS35 inhibitor provided by the invention targets host factors, is more difficult to induce Zika virus drug resistance, and has more lasting treatment potential.

Disclosed herein are prokaryotic homologs of viperin (pVips), and nucleotide and nucleoside analogs produced from pVips. These nucleotide and nucleoside analogs stop nucleotide chain synthesis and provide host cells with resistance to viral infections by targeting actively replicating viral genome. Further, these nucleotide and nucleoside analogs decrease DNA replication in malignant cells. Further disclosed are methods of identifying pVips, and nucleotide and nucleoside analogs produced thereof.

The invention relates to the field of cell engineering and gene engineering, in particular to a method for enhancing the multiplication capacity of umbilical cord mesenchymal stem cells and application. The expression or activity of the STN1 gene in the umbilical cord mesenchymal stem cells is reduced through an accelerant, and the umbilical cord mesenchymal stem cells with enhanced multiplication capacity are obtained. By knocking down the hUC-MSCs of STN1, the proliferation speed of the hUC-MSCs is higher, DNA replication is more active, the clone formation ability is higher, and the aging proportion of the hUC-MSCs can be remarkably reduced.

The present invention relates to cells comprising an orthogonal DNA replication machine, linear plasmids replicable by said machine, uses of said cells and said linear plasmids, methods of maintaining linear plasmids in cells, methods of evolving sequences of interest, and methods of preparing polypeptides or nucleic acids. The invention also relates to error-prone DNA polymerases, nucleic acids encoding said polymerases and uses thereof.

A non-integrating viral delivery system is disclosed. The system includes a viral carrier, wherein the viral carrier contains a defective integrase gene; a heterologous viral episomal origin of replication; a sequence encoding at least one initiator protein specific for the heterologous viral episomal origin of replication, wherein expression of the sequence encoding the at least one initiator protein specific for the heterologous viral episomal origin of DNA replication is inducible; and at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest.

The invention belongs to the field of new application of medicines, and particularly relates to application of 4-hydroxyderrisin in preparation of anti-influenza virus medicines. It is found for the first time that 4-hydroxyderrisin can significantly inhibit replication and internalization of influenza A virus H1N1 and has significant anti-influenza virus activity, the median inhibitory concentrations of 4-hydroxyderrisin in MDCK cells and A549 cells are 0.60 M and 0.23 M respectively, and the 4-hydroxyderrisin is low in cytotoxicity and high in safety. Experiments show that 4-hydroxyderrisin plays a role in the early stage of a virus life replication cycle, and plays an antiviral role by inhibiting virus internalization without affecting virus adsorption and neuraminidase activity. The medicine can be prepared into a solid or solution dosage form, is used for preventing and treating influenza, and has a good application prospect.

An agent that generates DNA replication stress and / or inhibits pathways involved in DNA replication stress tolerance for use in a method of treating a patient with cancer. The treatment comprises: determining whether the cancer expresses HORMAD1; and, if so, administering to said patient an agent that generates DNA replication stress and / or inhibits pathways involved in DNA replication stress tolerance.

The curcumin-coupled copper-gold bimetallic nanoszyme AuCu-Cur prepared by the application can remove ROS and relieve oxidative stress, prevent further damage to DNA, and introduce human umbilical cord mesenchymal stem cell-derived extracellular vesicles (hUCMSC-EVs) to improve the bioavailability. In the system, curcumin can repair damaged DNA by promoting the expression of NEIL3. In addition, the vesicles themselves are rich in various tissue repair-related proteins and RNAs, which can promote DNA replication of notochordal cells (NPCs), reverse the aging state and stimulate extracellular matrix synthesis. In the IVDD model induced by rat tail vertebra puncture, we confirmed that the material has strong antioxidant and DNA repair capacity and can effectively delay cell aging, thereby relieving the progression of IVDD. Compared with existing intervertebral disc biomaterial treatments, the present scheme can realize continuous DNA repair, which is an active, multi-target repair program that not only interrupts the vicious cycle of intervertebral disc degeneration but also promotes tissue transformation to regeneration.

The invention discloses a novel application of ginkgo biflavone in lung cancer treatment, belongs to the technical field of medicines, and researches show that ginkgo biflavone can target CDC7 and is a novel targeted medicine. It is confirmed for the first time that ginkgo biflavone regulates and controls the thermal stability and enzymatic degradation of ginkgo biflavone through targeting CDC7 kinase (binding free energies-8.3 kcal / mol), directly intervenes in the DNA replication process, and provides a natural drug lead compound for development of a novel CDC7 small-molecule inhibitor. The compound has great significance in preventing and treating malignant tumors, and has a wide prospect in the aspect of preparing medicines for treating lung tumors.

A comprehensive teaching aid system for genetic science at least including numerous (deoxy-) ribonucleotide models that can be assembled to form DNA / RNA single strands, or to form the beautiful DNA double helix structure when numerous adjacently and oppositely connected deoxynucleotide models are attached by magnets, tRNAs, and three different plates for DNA replication, mRNA transcription and protein synthesis respectively. The (deoxy-) nucleotide model includes a phosphate model, a (deoxy-) ribose model and a base model connected in sequence. Between two adjacently disposed (deoxy-) ribonucleotide models, a (deoxy-) ribose model is connected to a phosphate model in the head-to-tail fashion to form a detachable and flexible chain structure. The base model is laterally connected to the (deoxy-) ribose model, and two base models in two oppositely disposed deoxynucleotide models are flexibly and complementarily attached.

The invention discloses a high-yield cell-free protein synthesis system based on a cell crude extract and application of the high-yield cell-free protein synthesis system. The system comprises an escherichia coli extract, a DNA template, a buffer solution, an amino acid mixture, a nucleoside triphosphate mixture, an ATP regeneration system and antibiotics, the antibiotics are selected from compounds having inhibitory activity on metabolic processes except ribosome functions in prokaryotic microorganisms, including but not limited to one or more combinations of interfering DNA replication, DNA transcription, cell wall synthesis, energy metabolism, lipid synthesis or other non-translational related metabolic pathways. According to the method disclosed by the invention, the escherichia coli crude extract is used as a bacterial chassis, and non-essential metabolic pathways in a CFPS system are selectively inhibited by introducing different types of antibiotics, so that the yield of protein synthesis is increased.

A method for detecting hematopoietic stem cells comprises the steps that a CD133 nucleic acid aptamer functionalized ITO electrode is used as a substrate, e-Y-Click labeling of hematopoietic stem cell surface protein is achieved through in-situ electrochemical activation of PTAD-N3, N3 groups combined on the surface and DBCO modified nucleic acid chains T are subjected to SPAAC reaction, and the hematopoietic stem cells are obtained. The method comprises the following steps of: triggering a self-replication cycle reaction of DNA (Deoxyribose Nucleic Acid), enriching electrochemical signal substances on the surfaces of the hematopoietic stem cells, qualitatively judging the hematopoietic stem cells by detecting electrochemical signals, and correspondingly judging the quantity of the hematopoietic stem cells contained in a sample to be detected according to the established relationship between the intensity of the electrochemical signals and the quantity of the hematopoietic stem cells, so as to realize quantitative detection.

Formulations and uses thereof are provided for supporting human natural defense against viral infections as well as providing treatment for viral infections susceptible to a lysosomotropic agent. Administration of a lysosomotropic agent, such as ammonium chloride (NH4Q), can militate against the uncoating of viruses within the lysosome of an infected cell and thereby minimize infection by viruses whose replication cycle relies upon an uncoating step in such a manner.

Certain embodiments of the invention provide a recombinant polypeptide comprising a T5 DNA polymerase amino acid sequence operably linked to a DNA helicase amino acid sequence, as well as methods of using such a recombinant polypeptide for DNA replication and / or mutagenesis. Certain embodiments of the invention provide a targeted artificial DNA replisome complex. Certain embodiments of the invention provide a targeted DNA mutagenesis system.

Compounds represented by Formula I in which A0-A2, R0-R6, L, Z and Q are defined herein. The compounds of Formula I are CHK 1 inhibitors. Therefore, the compounds can be used to treat diseases, disorders and conditions associated with continuous activation of CHK1 or with high internal DNA damage or injury during DNA replication, such as cancers.

Formulations and uses thereof are provided for supporting human natural defense against viral infections as well as providing treatment for viral infections susceptible to a lysosomotropic agent. Administration of a lysosomotropic agent, such as ammonium chloride (NH4Cl), can militate against the uncoating of viruses within the lysosome of an infected cell and thereby minimize infection by viruses whose replication cycle relies upon an uncoating step in such a manner.

The application relates to the technical field of antibacterial peptides, in particular to application of an antibacterial peptide with antifungal activity, and discloses antifungal activity, antibiofilm activity and an action mechanism of Jelleine-Ic on Candida albicans. Jelleine-Ic shows very strong antifungal activity on Candida albicans. Jelleine-Ic destroys the integrity of the cell wall of Candida albicans and enhances the permeability of the cell membrane of Candida albicans. In addition, Jelleine-Ic interacts with DNA when entering the fungal cell, influences the expression of genes involved in DNA replication and repair, and induces the generation of active oxygen in the cell, thereby accelerating the death of the fungal cell. According to the above results, Jelleine-Ic can effectively inhibit the growth of Candida albicans and has the potential for treating fungal infections.

The present invention provides engineered T7 replisomes that contain one or more modified protein components, and E. coli based orthogonal DNA replication systems that contain such engineered T7 replisomes. Relative to known orthogonal replication systems, the engineered DNA replication systems of the invention are capable of evolving target polynucleotide sequence with enhanced mutation rates.

Gene amplifications are an oncogenic driver utilized by many forms of cancer in tumor development or treatment relapse. Promoting genomic instability or the proclivity to propagate genomic alterations through acquired defects in DNA repair machinery, replication licensing, or cell cycle control, gene amplifications not only drive oncogenesis, but also afford an opportunity for therapeutic exploitation. Here, CRISPR-Cas9 nickases are disclosed which selectively promote cancer cell death in a gene amplification-dependent manner. For example, CRISPR- Cas9 nickases generate a lethal number of highly toxic single-ended double-strand breaks within the genome of proliferating cancer cells harboring amplified genomic loci during DNA replication. Cas9 nickases may serve as a tumor-selective therapeutic that mitigates the collateral damage observed with conventional chemoradiotherapies and avoids chemoresistance.

The present invention discloses the mutation sites of SUV420H1 and its application in regulating the proliferation of tumor cells. The present invention provides the application of a substance capable of simultaneously site-directed mutating the K219 and R220 sites of the SUV420H1 protein in an organism or cell in any one of the following: inhibiting the proliferation of tumor cells; treating tumors, inhibiting DNA replication in tumor cells; reducing the binding of the SUV420H1 protein and the H2A.Z protein or its nucleosome. The experiments of the present invention prove that the K219R220 site on the SUV420H1 protein, which can affect the binding to H2A.Z, and mutating the K219R220 site on the SUV420H1 protein can inhibit the proliferation of tumor cells, indicating that the K219R220 site on the SUV420H1 protein can be used as a target to study specific drugs for inhibiting the proliferation of tumor cells and has potential clinical application value.

The invention relates to cells, such as bacterial cells, comprising orthogonal DNA replication machinery, linear plasmids that may be replicated by said machinery, uses of said cells and said linear plasmids, methods of maintaining linear plasmids in cells, methods of evolving sequences of interest, and methods of making polypeptides or nucleic acids. Orthogonal systems that are based on or derived from phage, such as phi29, that infect gram-positive bacteria are disclosed herein.

The invention discloses application of 5-fluorodeoxycytidine and a medicine, relates to the technical field of biological medicines, and particularly provides application of 5-fluorodeoxycytidine in preparation of a medicine for resisting staphylococcus aureus. The 5-fluorodeoxycytidine can act on staphylococcus aureus to replicate an initiator protein DnaA, inhibit the initiator protein DnaA and cause DNA replication disorder of the staphylococcus aureus, so that the activity of the staphylococcus aureus is effectively inhibited, and a bactericidal effect is achieved.

Mismatch anchoring compounds (MACs) are disclosed that recognize and bind to specific base-pair mismatches (mmBP) present within single stranded or double stranded DNA, RNA, and oligonucleotide sequences, and enable the ex vivo identification of DNA / RNA point mutations (DNAPM / RNAPM), including disease-associated, rare, and low abundance DNAPM / RNAPM, and the isolation and elimination of mmBP-positive cells. The use of MACs in vitro and in vivo (a) blocks mmBP-positive DNA replication and gene transcription / expression, (b) inhibits mmBP-positive cell proliferation, (c) reverses, prevents, and / or treats mmBP-rnediated disease, aging, and age-related disorders, (d) blocks mmBP-positive RNA and / or RNA-DNA duplex translation and inhibits the production of abnormal disease-causing proteins, (e) silence mmBP-positive genes, and (f) blocks intracellular pathogen replication. MAC-conjugates and their uses are disclosed, including MACs radiolabeled with SPECT / PET / particle-emitting isotopes for radioimaging and / or radiotherapy of mmBP-positive diseases.

The invention discloses an application of mPGES-2 as a target spot in preparation of a medicine for preventing and / or treating liver cancer. The invention proposes that mPGES-2 is a drug target for preventing and / or treating liver cancer for the first time. Experiments show that mPGES-2 knock-down significantly down-regulates expression of a cell proliferation antigen Ki67 and DNA replication marker protein PCNA, inhibits the S-phase process of cells, and further inhibits proliferation slowing caused by DNA replication of liver cancer cells; overexpression of mPGES-2 can obviously promote expression up-regulation of Ki67 and PCNA, promote the S-phase process of cells, enhance the DNA replication efficiency and finally cause obvious increase of cell proliferation; the mPGES-2 can regulate the proliferation of the liver cancer cells, so that the mPGES-2 can be used as a drug target for preventing and / or treating the liver cancer.

The invention is suitable for the technical field of preservative production, and provides a starch food preservative and a preparation method thereof.The preservative comprises the following raw materials: a dynamic covalent bond gel carrier, a polyphenol-metal nano-cluster compound, compound enzyme, a compound extract, modified zein and a multistage pore channel starch-based adsorbent, a dynamic covalent bond gel carrier is added, dynamic imine bonds are cross-linked to form a three-dimensional network structure, when the pH of putrefying bacteria produced acid is reduced, the imine bonds are broken, antibacterial components are released, cross-linking is carried out again after the pH rises, waste of active components is prevented, a double-pH response network is formed with modified zein, and a release curve is smoother; by adding a polyphenol-zinc nano-cluster compound, the microbial enzyme activity and DNA replication are inhibited; fe2O3 in the modified zein is subjected to photo-thermal response, is heated under illumination, softens the shell to release active components, and is linked with the dynamic gel carrier to realize chemical and physical double-trigger release.

The present invention relates to a technique for replicating polynucleotides within eukaryotic cells and transferring the polynucleotides between eukaryotic cells, where the polynucleotides include polynucleotide sequences encoding pores that secrete DNA and genes encoding proteins required for DNA replication. The polynucleotides may also include polynucleotide sequences that provide a desired function, such as a therapeutic effect, or polynucleotide sequences that can complement or directly replace mutated genes in eukaryotic cells. Treatment methods involving administration of the polynucleotides are also disclosed.

The present invention relates to a bacterial continuous evolution system, an orthogonal DNA polymerase and a continuous evolution method. The present invention combines an orthogonal DNA replication system with an orthogonal DNA polymerase to obtain a continuous evolution method that can contain all mutation types, can achieve mutations in long DNA fragments, has good continuity and is easy to operate. By inducing the opening and closing of DNA polymerase expression, the switching between the linear plasmid error-prone mutation process and the high-fidelity replication process is achieved, thereby achieving efficient continuous evolution of the target DNA sequence.

The invention discloses application of a host factor RAB10 in regulation and control of hepatitis B virus antigen expression and anti-hepatitis B virus. The invention provides an application of a host factor RAB10 or a coding gene thereof in regulation and control of hepatitis B virus replication or hepatitis B virus antigen expression level in host cells, and an application of the host factor RAB10 or an expression promoter thereof in preparation of drugs for resisting hepatitis B virus or inhibiting hepatitis B virus antigen. The invention also discloses application of the host factor RAB10 or the coding gene thereof as a drug target in screening drugs for resisting hepatitis B virus or inhibiting hepatitis B virus antigen. Researches show that RAB10 can significantly inhibit HBsAg expression, HBV DNA replication, HBV RNA transcription and HBs protein level, is expected to be used for developing a novel therapeutic drug for inhibiting hepatitis B virus surface antigen, and provides a new strategy for HBV functional cure.

This disclosure concerns compositions and methods for replicating DNA containing a T7 origin of replication (T7 or / ). Compositions and methods herein use a set of proteins that includes a DNA polymerase (DNAP), an RNA polymerase (RNAP), a single-strand binding protein (SSBP) and a helicase-primase (HP), along with one or more linearising proteins and linearising protein-binding nucleic acid sequences to maintain the replicated DNA in a linear conformation. In one example, the addition of a protelomerase recognition sequence (TeIRL) as the linearising protein-binding nucleic acid sequence to a T7 or / plasmid, in combination with the use of a protelomerase (TelN) as the linearising protein during DNA replication, enabled linearisation, replication and maintenance of a T7 or / plasmid in E. coli. The DNAP may be a mutagenic polymerase, so that methods herein may also be used for DNA mutagenesis and evolution of target DNA sequences.

The invention discloses a whole genome continuous mutation system based on base deaminase and application. The whole genome continuous mutation system comprises a MutaGB-A system and a MutaGB-B system. The MutaGB-A system is a series of plasmids for coding'DNA replication protein-base deaminase 'fusion protein, and whole genome continuous mutation can be formed after report strains are converted; the MutaGB-B system is a series of plasmids for coding'DNA replication protein-base deaminase-phase separation element 'fusion protein, after a strain containing'DNA replication / repair protein-phase separation element' fusion protein is converted, whole genome continuous mutation can be more efficiently introduced, and the mutation type is expanded to cover all SNP (Single Nucleotide Polymorphism) types. The whole genome continuous mutation system can improve the mutation rate of yeast strains by more than ten thousand times, is controllable and continuous, and is suitable for different biological species. By combining with a corresponding screening method, the method can be used for rapidly obtaining a target strain with specific characters.