22 results about "DNA Binding Motif" patented technology

Constructs comprising transactivation domains (TADs) for activating gene expression are disclosed herein, along with methods of using the same. Nucleotides encoding the same and vectors for the delivery of the nucleotides are also disclosed. In particular, synthetic transcription factors comprising a TAD fused to a heterologous DNA binding domain, and methods of using thereof are provided.

The present invention provides a method of identifying the amino acid position of a DNA modifying enzyme for insertion into a heterologous DNA binding domain (DBD), the method comprising the steps of: providing a library of DNA modifying enzymes wherein members of the library comprise heterologous amino acid sequence insertion throughout the DNA modifying enzyme; identifying those DNA modifying enzymes in the library that have DNA modifying activity; and determining insertion positions in those DNA modifying enzymes identified in the previous step. The invention also relates to a method of preparing a DNA modifying enzyme comprising an inserted heterologous DBD, a method of modifying a nucleic acid sequence in a cell, and a method of evolving a DNA binding domain for a desired target sequence. The invention also provides nucleic acid sequences encoding such DNA modifying enzymes and DNA binding domains, corresponding vectors and host cells.

The present invention relates to chimeric receptors that can be used in whole-cell sensors for detecting analytes of interest. The inventors showed that the DNA binding domains and downstream gene expression can be activated via dimerization of an artificial dimerization composed of a single chain variable domain. They demonstrated for the first time that an artificial bacterial receptor using an antibody-like domain can be activated and produce a transcriptional output upon ligand-binding. In particular, the present invention relates to a chimeric receptor polypeptide comprising: i) a first DNA binding domain, ii) at least one binding domain selected from the group consisting of heavy chain variable domain, camelid VHHs, or antibody mimetics having specificity for an analyte, and iii) a linker between the DNA binding domain and the binding domain.

The disclosure pertains to methods and compositions for the rapid and efficient transformation of plants. The disclosure further provides methods for producing a transgenic plant, comprising (a) transforming a cell of an explant with an expression construct comprising (i) a nucleotide sequence encoding a WUS / WOX homeobox polypeptide; (ii) a nucleotide sequence encoding a polypeptide comprising two AP2-DNA binding domains; or (iii) a combination of (i) and (ii); and (b) allowing expression of the polypeptide of (a) in each transformed cell to form a regenerable plant structure in the absence of exogenous cytokinin, wherein no callus is formed; and (c) germinating the regenerable plant structure to form the transgenic plant. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present disclosure.

The application discloses a mutant of a photosensitive protein BphP1. The application also provides a kind of optogenetic system, which comprises the fusion protein of the BphP1 mutant and the transcription activation domain (AD) of Gal4, the fusion protein of the photosensitive protein PpsR2 and the DNA binding domain (DBD) of Gal4, and the expression vector for expressing target genes through Gal1 promoter, wherein the PpsR2 can be wild type truncated variant Q-PpsR2. The induction intensity of the optimized optogenetic system of the application is 2.99 times that of the original light induction system.

The present invention is inter alia directed to a fusion protein comprising (i) an adenosine deaminase 2 (ADA2) domain and (ii) a DNA binding domain, wherein the DNA binding domain is capable of binding to a target DNA sequence; a pharmaceutical composition comprising adenosine deaminase 2 (ADA2) or a functional variant thereof and at least one pharmaceutically suitable excipient, wherein the pharmaceutical composition is targeted to the lysosome; and a pharmaceutical composition comprising adenosine deaminase 2 (ADA2) or a functional variant thereof and at least one pharmaceutically suitable excipient, wherein the pharmaceutical composition is targeted to a macrophage.

The invention discloses a GAL4-TEAD4 fusion protein, a YAP activation level detection kit and application of the GAL4-TEAD4 fusion protein and the YAP activation level detection kit, the GAL4-TEAD4 fusion protein comprises a GAL4 DNA binding structural domain and a TEAD4 YAP binding structural domain, and endogenous interference is eliminated by deleting the DNA binding structural domain of TEAD4; the kit comprises a GAL4-TEAD4 expression plasmid, a UAS-luciferase reporter plasmid and a luciferase detection reagent, and during detection, mammalian cells are co-transfected, a GAL4-UAS reporter system is utilized, and the YAP activity is directly quantified through a luciferase activity value. The system is high in sensitivity, can accurately capture activity change caused by YAP overexpression or siRNA knock-down, and is suitable for YAP activity detection and screening of small molecule compounds targeting YAP-TEAD interaction.

Provided are chimeric high mobility group (HMG) transcription factor polypeptides. In some aspects, provided are chimeric polypeptides comprising an N-terminal effector domain of a first HMG transcription factor and an HMG DNA binding domain and a C-terminal effector domain, wherein the HMG DNA binding domain or C-terminal effector domain is of a second HMG transcription factor. Also provided are cells (e.g., immune effector cells) that express a chimeric HMG transcription factor polypeptide of the present disclosure. In some instances, such cells (e.g., T cells) exhibit enhanced function relative to cells expressing only native wild-type HMG transcription factors. Also provided are methods of using such cells to treat a condition (e.g., cancer, infection, and / or the like) in a subject in need thereof.

The invention discloses a transcriptional activation type RUBY visual virus infection system as well as a method and application thereof. The system is based on a betacyanin biosynthesis pathway, a transcriptional activation system is utilized, and a yeast GAL4 DNA binding domain (BD) and a herpes virus VP16 transcriptional activation domain (AD) are fused and constructed in a virus vector; when the virus infects expressed transgenic tobacco, the expression of the RUBY whole gene can be activated, so that a red signal is generated. The construction method of the system is clear, complex instruments are not needed, an efficient and convenient tool platform is provided for protein function analysis, host factor screening and infection mechanism exploration in virology research, and good expandability is achieved.

Provided is a retrotransposase which comprises a target DNA binding domain comprising a zinc finger binding motif, a reverse transcriptase domain, and an endonuclease domain, and which is capable of reverse transcription of RNA to DNA. The invention also relates to a system for modifying DNA. The system comprises the retrotransposase or a nucleic acid encoding the retrotransposase; and a donor RNA or a nucleic acid encoding the donor RNA, the donor RNA comprising: a sequence that binds to the retrotransposase and a heterologous sequence.

Provided are designed proteins, derived from OCT4 and SOX2, which have enhanced stability and binding affinity for one another compared to the wild-type equivalents. These designed proteins are species-agnostic. Specifically provided are isolated SOX2 proteins having an artificial SOX2 DNA binding domain and isolated OCT4 proteins having an artificial OCT4 DNA binding domain. Also provided are uses of these proteins to generate induced pluripotent stem cells, vectors, and host cells containing these proteins.

This invention provides a technology that can improve the proliferation rate of cells in cultures using microcarriers. [Solution] The cell line is established by introducing the following four genes: CDK4(R24C), TERT, CCND1, and p53 lacking the DNA binding domain into bovine progenitor cells.

The present invention relates to methods and compositions for modifying a target site in the genome of a cell. Fusion proteins including one or more DNA binding domains and one or more heterologous domains, such as DNA modifying domains, connected by improved linker sequences are provided. Codon optimized polynucleotides encoding fusion proteins including one or more DNA binding domains and one or more heterologous domains connected by improved linker sequences are provided.

The application of mitoxantrone in inducing IRF4 protein degradation through autophagy-lysosome pathway belongs to the field of biological medicine. Mitoxantrone can specifically bind to IRF4 protein and significantly reduce the level of IRF4 protein. Mitoxantrone mainly binds to the DNA binding domain of IRF4, shortens the half-life of IRF4 protein, and promotes its degradation through the autophagy-lysosome pathway rather than the proteasome pathway. Mitoxantrone can interact with IRF4 protein and autophagy receptor p62 at the same time, promote the formation of IRF4 and p62 complex, enhance the ubiquitination modification of IRF4 and recruit it to the autophagy degradation pathway, thereby playing the role of molecular glue. Mitoxantrone treatment can inhibit the transcriptional activation function mediated by IRF4, down-regulate the expression of its downstream target genes, inhibit the proliferation of multiple myeloma cells and tumor growth, and provide a new technical approach for the development of anti-tumor drugs targeting transcription factors.

The application provides a Bst DNA polymerase domain, a mutant and application thereof, and belongs to the technical field of molecular biology. The amino acid sequence of the Bst DNA polymerase domain is shown in SEQ ID NO:1. The Bst DNA polymerase mutant is obtained by replacing the natural DNA binding domain of the wild-type Bst DNA polymerase with the amino acid sequence shown in SEQ ID NO:1. The Bst DNA polymerase mutant exhibits extremely high catalytic efficiency on dUTP, and also has excellent hot-start characteristics, and can reduce the false positive rate of LAMP technology to close to zero. The Bst DNA polymerase mutant also has extremely strong complex sample tolerance, and the detection sensitivity of the pathogen is 20 times higher than that of the wild-type Bst enzyme, and can meet the early and accurate diagnosis requirements of trace infection.

The invention is based on designed ankyrin repeat proteins (DARPins) specifically binding to a DNA binding domain of human p53. The DARPins of the invention are capable of forming a binding interface with the DNA binding domain of p53, and upon binding facilitate p53 protein stability. The DARPins of the invention are for use in the treatment of p53 related diseases such as cancer. Further, the invention pertains to nucleic acid constructs encoding the DARPin of the invention, methods for their production and cells comprising the DARPins or nucleic acid constructs of the invention.

The invention relates to a CAR-gamma delta T cell of a self-secreting immune cell adapter and a preparation method of the CAR-gamma delta T cell. The CAR-gamma delta T cell is prepared from a gamma delta T cell, the gamma delta T cell contains a nucleotide sequence for coding a SynNotch receptor and a nucleotide sequence for coding an effector, the SynNotch receptor comprises a transmembrane core module and an intracellular regulation and control module, the intracellular regulation and control module is connected with the C end of the transmembrane core module, and the C end of the intracellular regulation and control module is connected with the C end of the transmembrane core module. The intracellular regulation module comprises a DNA binding domain and a transcriptional regulation structural domain, and the transmembrane core module can be cut by gamma-secretase to separate the intracellular regulation module; the nucleotide sequence encoding the effector includes an upstream activation sequence capable of specifically binding to the DNA binding domain, and nucleotide fragments encoding an immune cell adapter, a self-cleaving peptide, and a T cell immune activator.

The invention provides a wheat gene-protein multi-modal information fusion and visualization method, relates to the technical field of protein structure prediction, designs a set of improved scheme combining artificial intelligence deep learning fusing geometric constraint and calculation optimization, and a constructed framework breaks through the limitation of traditional single-modal analysis. End-to-end generation from wheat genome regulation information to a protein three-dimensional structure is realized; according to the method, a subgenome specific noise scheduling mechanism is introduced into diffusion model training, folding preference of different subgenomes can be effectively distinguished, limitation of traditional sequence alignment is broken through, alpha-helix stability change of a DNA binding domain is successfully predicted, a dynamic mask mechanism is developed, a specific regulation element is allowed to be shielded in the generation process, and the method is suitable for large-scale popularization and application. Therefore, directional repair of the functional protein scaffold is realized, a dynamic visualization system is established, a time dimension is introduced, and conformation evolution of a wheat protein structure in different development stages or under different stress conditions can be dynamically presented.

Modified filamentous fungi with improved protein productivity, and proteins using the modified filamentous fungi. Providing a manufacturing method. [Solution] An ACE3 variant is expressed, and the ACE3 variant is Zn(II)2C of ACE3. A modified filamentous fungus in which substantially all of the ys6 type DNA binding domain is missing. A method for producing proteins, including culturing filamentous fungi.

The present disclosure provides DNA binding proteins engineered to bind selectively to engineered synthetic enhancer elements that are orthogonal to a host genome. Introduction of the DNA binding protein and the synthetic enhancer is used, in several embodiments, to modulate gene expression of a gene of interest without disruption or alteration of other genes within the host cell genome. In several embodiments, the DNA binding protein selected is a zinc finger protein.

To provide pharmaceutical compositions for synergistically modulating microtubule associated protein tau (MAPT) gene expression in a cell.SOLUTION: A pharmaceutical composition comprises: a first genetic modulator which comprises a first DNA-binding domain that binds to a first target site of at least 12 nucleotides in the MAPT gene, and a first transcriptional regulatory domain or a first nuclease domain; and a second genetic modulator which comprises a second DNA-binding domain that binds to a second target site of at least 12 nucleotides in the MAPT gene, and a second transcriptional regulatory domain or a second nuclease domain; where the binding of the first genetic modulator to the first target site and the binding of the second genetic modulator to the second target site results in synergistic repression of MAPT gene expression.SELECTED DRAWING: Figure 13

A viral gene therapy vector for removing hepatitis B virus and application, relates to the field of biological medicine, especially to a gene therapy vector and its preparation method and application. To solve the problem that the existing viral gene therapy vector can only inhibit hepatitis B virus replication and cannot remove viral cccDNA. The vector comprises a promoter, an intron, an enhancer, a PGLYRP2 protein nuclear localization signal NLS functional domain coding gene, a PGLYRP2 protein HBV DNA binding domain coding gene, a PGLYRP2 protein secretion signal peptide functional domain coding gene, a PGLYRP2 protein HBV nuclear envelope binding functional domain coding gene and an IRES fragment. The viral gene therapy vector of the application can significantly promote the removal of HBV in the liver of mice. The viral gene therapy vector of the application is used for treating hepatitis B virus infection.