19 results about "Integrase" patented technology

Among other things, provided herein are systems that replace the natural random integration activity of a retrovirus with site-specific integration machinery. This approach allows for a more precise targeting of a gene of interest into a human genome, e.g., for therapeutic purposes. The system may include integration-deficient retrovirus (e.g., lentivirus) (IDLV), in which the natural integration activity has been reduced (e.g., by mutation to the viral integrase polypeptide). Instead, the system may comprise a site-specific recombinase (e.g., a serine recombinase, e.g., a serine integrase) capable of directing insertion of a template DNA, or portion thereof, into a desired site in the human genome.

The invention discloses a mycobacteriophage and application thereof, and relates to the technical field of bacteriophages. The mycobacteriophage is named as Mycobacter phage BZNK001, is preserved in Guangdong Microbial Culture Collection Center, has a preservation number of GDMCC No: 67907-B1, does not contain antibiotic drug-resistant genes and integrase genes, has excellent biological safety and good environmental tolerance, can maintain high biological activity in a temperature range of 37-50 DEG C under an acid-base condition of pH 5-9, and can be used for preparing the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage for preventing and treating the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage for preventing and treating the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage. The bacillus subtilis has broad-spectrum and high-efficiency non-tuberculous mycobacterium cracking activity, and can effectively crack four clinical key mycobacteria such as mycobacterium abscessus, mycobacterium avium, mycobacterium intracellularis and mycobacterium smegmatis. According to the invention, a mycobacteriophage resource library is enriched, a novel and stable biological prevention and control means is provided for efficient prevention and control of mycobacteria, and the mycobacteriophage has good clinical application and industrialization values.

The invention provides a method for synchronously and rapidly detecting Escherichia coli and multiple drug-resistant genes thereof, which comprises the following steps: culturing a sample at night to obtain a bacterial solution to be detected, adding a buffer solution and a metal bath, centrifuging, taking a supernatant, detecting by micro-fluidic, optimizing the extraction process of nucleic acid in the sample, designing an optimal RPA primer probe group, and detecting the multiple drug-resistant genes of the Escherichia coli. A reasonable RPA coupled microfluidic detection system is established, synchronous and rapid detection of the escherichia coli uidA and I-type integrase gene intI1 thereof and drug-resistant genes sul1 and aadA5 is realized, the specificity is high, the sensitivity is high, the reaction time is short, the operation is convenient, and a method support is provided for monitoring and prevention and control of food-borne drug-resistant bacteria.

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.

The invention discloses a test strip biosensor for detecting uranyl ions, the test strip biosensor comprises a reaction system and a test strip, the reaction system comprises a DNAzyme substrate chain and a DNAzyme enzyme chain; the test strip comprises a substrate, a sample pad, a coupling pad, a nitrocellulose membrane and an absorption pad, the sample pad, the coupling pad, the nitrocellulose membrane and the absorption pad are sequentially overlapped on the substrate, a gold nanoparticle-DNA probe I compound is fixed on the coupling pad, a test line and a control line are arranged on the nitrocellulose membrane, an SA-biotin-DNA probe II is fixed on the test line, and an SA-biotin-DNA probe II is fixed on the control line. And an SA-biotin-DNA probe III is fixed on the control line. Compared with the prior art, the biosensor can realize high-sensitivity and high-specificity detection of uranyl ions by integrating a DNA enzyme induced cleavage reaction and a gold nanoparticle mediated colorimetric detection technology, the detection process is simple and rapid, and an important technical support is provided for field monitoring of UO2 < 2 + > in environment and biological samples.

The present disclosure relates to the field of biotechnology, and particularly relates to detection of BXB1 integrase. The present disclosure establishes a method for detecting BXB1 integrase based on double-antibody sandwich ELISA technology, which has good sensitivity and specificity, and the method is simple to operate and can quickly realize accurate quantification of BXB1 integrase; in addition, the present disclosure also provides an ELISA kit for detecting BXB1 integrase. By using the BXB1 integrase detection method or kit provided by the present disclosure, accurate quantification of the residual amount of BXB1 integrase in various intermediate products, semi-finished products or finished products in the process of preparing recombinant biological products by the BXB1 integrase system can be realized, which is beneficial to the quality control of products by biopharmaceutical enterprises.

This invention discloses a biosensor for detecting uranyl ions using a test strip, comprising a reaction system and a test strip. The reaction system includes a DNAzyme substrate chain and a DNAzyme enzyme chain. The test strip includes a substrate and a sample pad, a coupling pad, a nitrocellulose membrane, and an absorbent pad sequentially overlapped on the substrate. A gold nanoparticle-DNA probe I complex is immobilized on the coupling pad. A test line and a control line are provided on the nitrocellulose membrane. An SA-biotin-DNA probe II is immobilized on the test line, and an SA-biotin-DNA probe III is immobilized on the control line. Compared with existing technologies, this biosensor integrates a DNase-induced cleavage reaction with gold nanoparticle-mediated colorimetric detection technology, achieving high sensitivity and high specificity detection of uranyl ions. Furthermore, the detection process is simple and rapid, providing a solution for detecting UO2 in environmental and biological samples. 2+ This provides important technical support for on-site monitoring.

The application discloses streptomyces 891-DC with high aurovertin A yield and application thereof. The application uses CRISPR-Cas9 technology to first change an incomplete attB site sequence in the genome of streptomyces 891 into a complete attB site sequence, promote a plasmid containing a target gene chry, an integrase phiC31 gene and an attP site, and improve the integration rate of the target gene through specific combination of the attP site and the attB site on the genome. Secondly, a promoter capable of obviously improving the yield is screened to express the aurovertin biosynthesis gene cluster, so that the more stable and higher-yield streptomyces 891-DC is obtained. The aurovertin A shake flask fermentation yield is 1411.23+ / -29 mg / L, which is 60% higher than that of the original strain, and the genetic stability is good, thereby laying a technical foundation for industrial production of aurovertin.

The present invention relates to a system comprising a landing pad, a delivery nucleic acid and an integrating enzyme allowing selection of a specific gene of interest. The invention further relates to cells comprising the system, a method for providing a gene of interest and selecting a cell with a gene of interest, a landing pad nucleic acid, a delivery nucleic acid and a kit for providing and selecting a cell with a gene of interest.

In the present invention, there is provided a nucleic acid molecule that is capable of recognizing and / or binding to HIV, in particular to the integrase (IN) of HIV-1 and HIV-2 variants or portions thereof. In particular, the nucleic acid molecule is capable of binding to the HCINp1 (11 aa) (SEQ ID NO 4, PYNPQSQGVVE), HCINp2 (9 aa) (SEQ ID NO 5, NFKRKGGIG), HCINp3 (9 aa) (SEQ ID NO 6, LLWKGEGAV), HCINp4 (8 aa) (SEQ ID NO 7, IKVVPRRK) and / or HCINp5 (10aa) (SEQ ID NO 15, LLWKGEGAVV). The nucleic acid molecule may be a polynucleotide or an oligonucleotide.

The application discloses a corynebacterium striatum phage and application thereof. The corynebacterium striatum phage is a corynebacterium striatum phage CSP1, the preservation number of which is CGMCC NO: 45598, and the preservation time is July 4, 2023. The corynebacterium striatum phage has strong tolerance, biological safety and carries site-specific recombination functional elements (integrase and recombination sites). Meanwhile, the corynebacterium striatum phage can lyse 20 strains of multi-drug resistant corynebacterium striatum, and lays a foundation for subsequent medical researches using the corynebacterium striatum phage as a test and treatment carrier, a targeted pathogenic bacterium or symbiotic bacterium, and an immune regulator inducer.

The invention provides methods for genetically engineering T cells in vivo comprising administering to the subject a cytokine or nucleic acid molecule encoding a cytokine to recruit the subject's T cells to the administration site; followed by administration of a nucleic acid molecule encoding an antigen receptor. In some instances, the method also includes administering an integrase or nucleic acid molecule encoding an integrase to integrate the sequence encoding the antigen receptor into the DNA of the recruited T cells.

The application discloses an enzymatic colorimetric encoding-decoding system and application for multiple biomarker weighted detection, and belongs to the field of biomedical detection. The enzymatic colorimetric encoding-decoding system can realize the mapping relationship between color and multidimensional biomarkers through direct visual interpretation or simple spectrum analysis. The application further provides an enzymatic colorimetric encoding-decoding method for multiple biomarker weighted detection. By integrating enzyme-catalyzed multicolor reaction, nucleic acid-driven signal amplification and optical encoding-decoding mechanism, the multidimensional biomarker information is converted into quantifiable and visually analyzable optical signals, realizing single-tube multidimensional joint detection, high information density output and rapid disease state discrimination. The application has been successfully applied to early screening of pancreatic cancer, and can be extended to dynamic monitoring of multiple markers of other cancers and infectious diseases in the future, providing an efficient and low-cost solution for precision medicine and digital medicine.

A herpes simplex virus (HSV) gene vaccine and expression vector and / or vaccine vector comprising an HSV genome comprising: a complete deletion of glycoprotein G-encoding gene, glycoprotein J-encoding gene, glycoprotein D-encoding gene, and C glycoprotein I-encoding gene; a heterologous nucleic acid comprising an expression cassette and inserted in a region of the genome from which the glycoprotein G-encoding gene, glycoprotein J-encoding gene, glycoprotein D-encoding gene, and glycoprotein I-encoding gene have been deleted; an attL sequence; and an attR sequence, wherein the attL sequence is adjacent to a first end of the expression cassette and the attR sequence is adjacent to a second end of the expression cassette, and wherein the expression cassette comprises in operable communication a promoter and at least one gene encoding a heterologous protein.

The present disclosure provides compositions comprising engineered integration enzymes / eLSR and methods of using the same. In certain embodiments, the engineered integration enzyme comprises mutation(s) that substantially maintain or enhance integration activity at a pair of cognate integration recognition sites, and substantially decrease off-target integration activity at a pair of off-target integration recognition sites, when compared to a corresponding large serine integrase without said one or more substitutions (cLSR). The eLSR may further comprise a stabilization domain that increases the stability of the integration enzyme as compared to integration enzymes not comprising the stabilization domain.

The invention provides a construction body for specifically inserting a DNA (Deoxyribonucleic Acid) fragment and a transgene site into a genome, wherein the construction body comprises recombinase. The enzyme includes a modified HIV-1 integrase (HIV IN) tetramer having a plurality of mutations to facilitate heterodimer construction of the heterodimer wherein a first outer monomer and a second outer monomer are bound to a first inner monomer and a second inner monomer, respectively. A transcriptional activator-like effector (TALE) binds to the N-terminal region of the outer monomer via a polypeptide linker. The two TALEs confer enzyme sequence specificity and prevent non-specific interactions with the target DNA structure. The modified HIV IN monomer ensures the binding of the TALE to the outermost layer monomer and the arrangement of the TALE relative to the catalytic region of the HIV IN. A polynucleotide is bound to an internal monomer of the HIV IN. Upon transport to the nucleus alone or within the lentiviral capsid, the TALE interacts with DNA to locate the HIV IN, which then integrates the polynucleotide.

The present invention relates to the field of gene transfer, in particular, targeted gene transfer and enzymes for use in targeted gene transfer. In particular, the present invention provides a nucleic acid encoding a fusion protein having serine integrase activity, wherein the fusion protein comprises a serine integrase consisting of a protein having at least 80% sequence identity to the large serine integrase Pa01 and a nuclear localization sequence at the C-terminus of the serine integrase. The invention further provides a fusion protein encoded by said nucleic acid as well as methods of preparing an engineered cell using the nucleic acids and / or fusion proteins of the invention. Finally, the present invention provides various pharmaceutical compositions and their use in treating a disease or disorder.

This disclosure provides nucleic acid constructs for use in enhancing the site-specific insertion of an exogenous nucleic acid into a genome. In some embodiments, the nucleic acid construct comprises a first polynucleotide sequence encoding a DNA-binding protein engineered to bind to a specific genomic DNA sequence, a second polynucleotide comprising a modified integrase or a modified transposase that enables insertion of the exogenous nucleic acid into the genome, and a nucleic acid sequence encoding a linker between the two nucleotides. In some embodiments, the nucleic acid construct encodes a fusion protein, for example, a fusion protein for delivery to a cell via a lentiviral particle.