96 results about "Modified dna" patented technology

The invention discloses a nano magnetic bead for specific capture of DNA (deoxyribonucleic acid) of bacteria and fungi, a preparation method of the nano magnetic bead and a rapid sterile detection method of a cell therapy product. The method comprises the following steps: S1, coating Fe3O4 with polydopamine to form a modified Fe3O4 magnetic bead; s2, compounding the MOF shell layer to the modified Fe3O4 magnetic beads to form Fe3O4-coated MOF magnetic beads; s3, gold nanoparticles are anchored to the Fe3O4 (at) MOF magnetic beads to form Fe3O4 (at) MOF (at) Au magnetic beads; and S4, modifying the Fe3O4 (at) MOF (at) Au magnetic bead with a specific capture probe aiming at DNA of bacteria and fungi to form a Fe3O4 (at) MOF (at) Au-Probe probe magnetic bead. The modified DNA probe disclosed by the invention can efficiently capture free bacteria and fungus nucleic acid fragments in a sample, so that template enrichment is realized.

The invention relates to the field of DNA nanotechnology, and discloses a DNA origami structure based on functionalized magnetic nanoparticles, a preparation method and application, and the preparation method comprises the following steps: step 1, constructing a DNA origami structure with a cavity through a self-assembly method; the short chains in the DNA origami structure comprise a first short chain, a second short chain and a third short chain; 2, connecting a sulfydryl-functionalized first sequence to the first short chain, and chelating ferrous ions on the first sequence to obtain a sulfydryl-modified DNA origami structure; 3, ferrous ions chelated on the sulfydryl modified DNA origami structure are reduced, and a magnetic functionalized DNA origami structure internally loaded with Fe2O3 particles is obtained; and 4, connecting an antibody to a second short chain in the DNA origami structure through base complementary pairing to obtain the required DNA origami structure. According to the invention, DNA origami is combined with magnetic nanoparticles, efficient and controllable biological magnetic beads are obtained through dual immobilization and antibody modification, and a traditional application scene of DNA origami is broken through.

The disclosure provides, for example, DNA molecules comprising at least one nucleotide with a macromolecule, small molecule, or reactive handle (e.g., click handle) appended to a phosphorothioate backbone. The disclosure also provides methods for making such DNA molecules, for example using click chemistry. In some embodiments, the DNA molecule comprises a sequence that encodes an effector (e.g., a therapeutic effector).

Provided in some aspects are epigenetic-modifying DNA-targeting systems, such as CRISPR-Cas / guide RNA systems, that bind to or target a target site in a gene or regulatory element thereof in a T cell. In some aspects, the provided epigenetic modifying DNA-targeting systems provided herein modulate a T cell phenotype or activity. In particular, the provided embodiments relate to the transcriptional repression of genes to promote a stem cell-like memory T (TSCM) cell phenotype. In some aspects, also provided are compositions, polynucleotides, vectors, cells, and pluralities and combinations thereof, and methods and uses related to the provided epigenetic-modifying DNA-targeting systems, for example in modulating the phenotype in T cells including in connection with adoptive T cell therapy.

The invention discloses a nuclear targeting peptide modified DNA nano-particle based on a microfluidic technology and a preparation method and application thereof, and relates to the technical field of biological medicine, a poly (beta-amino ester) cationic polymer is used as a gene delivery carrier, the gene delivery carrier comprises a microtubule related sequence MTAS and a nuclear localization signal NLS oligopeptide, the gene delivery carrier and the plasmid DNA carrying the target gene are mixed to prepare nano-particles, and the plasmid DNA carrying the target gene is uniformly dispersed in the nano-particles by adopting a microfluidic mixing process; the nuclear targeting peptide modified DNA nano-particle is ingenious in structural design, the mixing uniformity of a nano-carrier and plasmid DNA is improved by adopting a micro-fluidic mixing process, the performance of the nano-particle is further improved by optimizing dialysis conditions, potential cytotoxicity and adverse effects are reduced, and the nuclear targeting peptide modified DNA nano-particle is suitable for large-scale production. And after transduction of non-activated T cells, strong tumor killing ability and cytokine secretion can be generated.

The invention provides a retrotransposase, which comprises a target DNA binding structural domain containing a zinc finger binding motif, a reverse transcriptase structural domain and an endonuclease structural domain, and can be used for reversely transcribing RNA (Ribonucleic Acid) into DNA. The amino acid sequence of the retrotransposase is shown as any one of SEQ ID No.1 to 6 or SEQ ID No.32 to 43 or SEQ ID No.68 to 71 or has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% and 99% identity with the amino acid sequence of any one of SEQ ID No.1 to 6 or SEQ ID No.32 to 43 or SEQ ID No.68 to 71. The invention also relates to a system for modifying DNA. The system comprises the retrotransposase provided by the invention or a nucleic acid encoding the retrotransposase provided by the invention; 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.

The present invention relates generally to compositions, systems and methods for modulating hepatitis B virus (HBV) by targeting gene repression. In particular, the present invention provides an epigenetically modified DNA targeting system, such as a CRISPR-Cas / guide RNA (gRNA) system, for transcriptional repression of hepatitis B virus genes to promote cell phenotypes that lead to reduced HBV infection. In some embodiments, the epigenetically modified DNA targeting systems of the present invention bind to or target a target site of at least one gene in the hepatitis B virus DNA sequence or a regulatory element thereof in a cell. In some aspects, the systems provided herein relate to transcriptional repression of one or more hepatitis B virus genes and / or regulatory elements thereof. In some aspects, the invention also provides methods and uses associated with the provided compositions, such as in repression of hepatitis B virus replication and expression associated with hepatitis B infection.

Disclosed are compositions, methods, and kits for modifying DNA within cells as well as compositions and methods for modifying gene expression in a cell. In particular, the invention generally relates to compositions, methods, and kits for DNA editing using single-stranded DNA. Compositions and methods for modifying gene expression using artificial microRNAs (amiRNA) are also contemplated.

Provided is a means for constructing a modified genomic DNA which has a nucleotide sequence different from an existing genomic DNA sequence, contains a modified gene capable of expressing a polypeptide, a functional RNA, or both, and does not contain exogenous DNA.　This method comprises: performing, at least twice, (a) a step for specifically cleaving genomic DNA of a cell into at least two target sequences by means of a sequence-specific nuclease, and linking the resultants through a DNA double-stranded cleavage repair by the cell; and deleting at least two regions from an existing gene and non-coding regions adjacent thereto to thereby produce a modified gene.

The invention provides an FSH33 peptide modified DNA tetrahedron nano delivery system as well as a preparation method and application thereof, and aims to provide a safe and efficient antioxidant nano drug and optimize the enrichment efficiency to promote ovarian aging and ovarian tissue damage repair. The system is a nano-scale nucleic acid compound composed of a tetrahedral DNA frame structure and an FSH targeting peptide. The tetrahedral DNA frame structure is a microscopic 3D stereoscopic tetrahedral structure formed by self-assembly of DNA single chains of a specific nucleic acid sequence, wherein each side extends out of a viscous end used for being connected with an FSH peptide. The research finds that the DNA tetrahedron can safely and effectively improve the ovarian endocrine function, ovarian reserve, fertility and the like of mice in a drug protection group, has no obvious toxic or side effect on all organs and tissues of the whole body, and can promote the repair of ovarian oxidative damage.

The invention provides a short polyarginine modified DNA probe as well as a preparation method and application thereof, and belongs to the technical field of biological analysis and detection. The preparation method of the short polyarginine modified DNA probe comprises the following steps: dissolving base-deficient site DNA in a buffer solution, and adding a coupling molecule linker for reaction to obtain an intermediate; and adding the purified intermediate into an Arg5-PEG1000-azide solution, and carrying out a click chemical reaction, so as to obtain the short polyarginine modified DNA probe. The invention also provides the short polyarginine modified DNA probe and application thereof. The short polyarginine modified DNA probe can generate characteristic and recognizable current signals through the nanopores so as to realize enzyme and small molecule detection, and has the advantages of high signal frequency, strong specificity, simple synthesis and great application potential.

Epigenetically modified DNA targeting systems, such as CRISPR-Cas / guide RNA (gRNA) systems, are provided that bind to or target one or more target sites in the IL-2 gene or its regulatory elements in cells such as lymphoid cells (e.g., T cells). In some embodiments, the provided epigenetically modified DNA targeting systems modulate the phenotype or activity of lymphoid cells, or the function of lymphoid cells such as the phenotype or activity of T cells. In some embodiments, methods and uses relating to the provided compositions in the modulation of lymphoid cells such as T cells are also provided, for example, in connection with adoptive T cell therapy.

We describe compositions of DNA donor molecules compatible with various gene insertion methods with major advantages for delivery, nuclear localization, and immune evasion. The modified DNA donor molecules are largely single stranded but contain short regions of double-strand DNA to reconstitute enzyme binding sites, and can be combined with other approaches to enhance donor recruitment into the nucleus.

This invention relates to DNA-functionalized thermosensitive phase transition polymers, their preparation methods, and applications, belonging to the interdisciplinary field of functional polymers and biomaterials. It aims to solve the technical problems of existing exosome separation methods, such as long processing time, low purity, high cost, and inability to specifically separate exosome subtypes. This invention provides a polymer prepared via free radical polymerization using N-isopropylacrylamide and acrylamide-modified DNA as monomers, and ammonium persulfate and tetramethylethylenediamine as initiators. This polymer exhibits reversible phase transition properties at physiological temperatures and is applied to the separation and detection of exosome subtypes. Specific capture and quantitative analysis are achieved through DNA recognition probes and fluorescently labeled antibodies. The polymer of this invention is simple to operate, quick to prepare, low in cost, and high in purity. It requires only a common centrifuge for non-destructive separation of exosomes and can integrate detection functions, providing technical support for diagnosis and detection.

The invention discloses a nano enzyme with catalyst inactivation resistance and a preparation method and application thereof, and the preparation method comprises the following steps: 1) performing functional pretreatment on DNA to obtain functional modified DNA for subsequent DNA coating construction; and 2) connecting a certain size of metal nanoparticles with catalytic activity with the functionalized modified DNA to construct the DNA coating-coated nano enzyme with catalyst inactivation resistance. The DNA coating on the surface of the prepared nano-enzyme can effectively prevent catalyst inactivation caused by particle aggregation in a high-salt environment, meanwhile, the coating has the function of a molecular sieve, can effectively inhibit non-specific adsorption of biomacromolecules such as protein and the like, allows a small-molecule substrate to be in contact with a catalytic interface, and improves the catalytic activity of the nano-enzyme. Therefore, catalytic activity is maintained in complex biological environments such as serum, sensitive detection of molecules is realized, and a solution is provided for precise medical diagnosis.

To provide means for constructing a modified DNA having a nucleotide sequence different from an existing genome DNA sequence, which is a modified genome DNA including a modified gene in which polypeptide, functional RNA or both can be expressed, but not including a foreign DNA.SOLUTION: A method includes (a) performing a step of cleaving a genome DNA of a cell in at least two target sequences more specifically by a sequence-specific nuclease, and connecting them by DNA double-strand break repair by a cell at least twice, deleting at least two regions from an existing gene and its adjacent non-encoded region to create a modified gene.SELECTED DRAWING: Figure 1A

Provided herein are methods for detecting the presence or progression of a disease in a subject as well as a method for determining the efficacy of a therapy for the disease by administering to the subject a probe comprising a phosphorothioated DNA barcode sequence, wherein the DNA barcode sequence comprises about 36 nucleotides to about 72 nucleotides, and wherein the DNA barcode is released when the disease is present in the subject; obtaining a biological sample containing the released DNA barcode from the subject; and amplifying and detecting the DNA barcode in the biological sample.

The invention relates to a double DNA nanosphere probe (DDNS) as well as a preparation method and biosensing application thereof. The DDNS comprises a DNA (deoxyribonucleic acid) nanosphere 1 (DNS-1) and a DNA nanosphere 2 (DNS-2). The DNS-1 and the DNS-2 are respectively formed by self-assembling two cholesterol modified DNA single-stranded probes with hairpin structures, and are respectively used for identifying target miRNA and triggering a non-enzymatic circulating strand displacement amplification (T-SDA) reaction. The DDNS can realize high-sensitivity fluorescence detection on target miRNA and can effectively inhibit non-target miRNA interference, and the interference inhibition rate is close to 100%. Besides, DDNS has excellent nuclease stability, the system can automatically enter living cells without a transfection reagent, high signal-to-noise ratio fluorescence differentiation of cancer cells and healthy cells is realized on the premise of maintaining cell activity, the performance is equivalent to that of a transfection agent dependent system, and the biological safety is remarkably improved. The invention provides a new material and a new method for application of the DNA nanostructure in the fields of molecular diagnosis and precision medical treatment.

The invention relates to the field of biological detection, in particular to a recombinase activity detection method and a kit. According to the method for detecting the strand displacement activity of the recombinase, the strand displacement template is subjected to fluorescence labeling, a quenching group modified DNA strand is displaced by utilizing the strand displacement effect of the recombinase, and the relative activity of the recombinase to be detected is obtained through a linear fitting equation of the fluorescence intensity increasing rate and the activity. The method can be used for detecting strand displacement products through a fluorescence detection means, is non-toxic and harmless, is simple and feasible, and can be used for determining the strand displacement recombination activity of the recombinase within 1 hour. By using the method, quantitative data of strand displacement activity can be obtained, and the strand displacement activity of recombinase can be accurately analyzed.

The invention relates to a perfluoroalkyl-modified DNA (deoxyribonucleic acid) nanostructure as well as a preparation method and application thereof. The method comprises the following steps: modifying a perfluoroalkyl chain on single-stranded DNA by using a solid-phase synthesis method to obtain perfluoroalkyl modified single-stranded DNA, and mixing the perfluoroalkyl modified single-stranded DNA according to a certain proportion by using a DNA dynamic self-assembly technology to prepare a perfluoroalkyl modified DNA tetrahedron, or preparing a perfluoroalkyl modified DNA tetrahedron by using a DNA origami technology to prepare a perfluoroalkyl modified DNA tetrahedron. The method comprises the following steps: designing and synthesizing a DNA nano structure with n hybrid functional arm chains, and hybridizing the DNA nano structure with perfluoroalkyl-modified single-stranded DNA to obtain the perfluoroalkyl-modified rectangular DNA origami. The method has the advantages of being simple in synthesis method, good in biocompatibility, diversified in function, high in loading capacity and the like, not only can provide excellent carriers for hydrophobic drug molecules and fluorescent small molecules, but also can be used as an ideal imaging molecular probe for < 19 > F-nuclear magnetic resonance. The method has a good application prospect in the fields of biosensing, drug delivery, antigen detection, molecular imaging, disease targeted therapy and the like.

The application discloses a kind of for sweat glucose concentration monitoring method, it is related to sweat monitoring field, comprising the following steps: step 1, enzyme-fixed chain conjugate synthesis: glucose dehydrogenase is reacted with bilirubin oxidase respectively with bifunctional crosslinking agent and is purified, obtain enzyme-crosslinking agent intermediate;It is clicked with azide modified DNA fixed chain chemistry reaction again, and after purification, obtain GDH-fixed chain and BOx-fixed chain conjugate;Step 2, preparation of circular DNA template: after annealing hybridization of phosphorylated template chain and primer chain, it is connected into ring by T4 DNA ligase;Step 3, rolling circle amplification self-assembly package enzyme DNA hydrogel;The present application uses super-flexible DNA hydrogel as flexible electrode material, realizes the efficient fixation and continuous stable catalysis of GDH and BOx, and constructs flexible sweat-based EBFC self-powered sensor as flexible bioelectrode, to realize continuous non-invasive blood glucose monitoring.

In various embodiments methods of utilizing gene drive constructs in asexual organisms such as viruses are provided. In certain embodiments the methods involve transfecting or infecting a cell with a modified DNA virus containing a gene drive construct; and infecting the cell with the target virus where the genome of said target DNA virus is modified by insertion of the gene drive construct into the genome of the target DNA virus and a population of modified target viruses (containing the gene drive construct) is produced.

The application belongs to the technical field of molecular biology, and discloses an acid anhydride modified DNA polymerase, a preparation method thereof, a PCR reaction reagent and application. The acid anhydride modified DNA polymerase is obtained by subjecting the DNA polymerase to an acid anhydride modification reaction with an acid anhydride compound, and the DNA polymerase comprises an amino acid fragment with a sequence shown in SEQ ID NO:1. The acid anhydride modified DNA polymerase has good anti-inhibition effect and can be well applied to extract-free PCR.

Methods for mitigation of methylation bias during preparation of sequencing libraries are described. The methods may comprise, e.g., extracting DNA fragments from a sample; performing at least one of: (i) a first end repair reaction, where the first end repair reaction comprises the use of a non-strand-displacing DNA polymerase; or (ii) a second end repair reaction, where the second end repair reaction comprises the use of a chain termination mechanism; performing at least one of: (iii) a tailing reaction to add an overhanging polynucleotide strand to the end-repaired DNA fragments, where the tailing reaction comprises the use of a single deoxynucleotide triphosphate (dNTP); or (iv) a nick / gap repair reaction using a DNA ligase to fill in single- stranded nicks or gaps in the end-repaired DNA fragments, to generate a plurality of modified DNA fragments; and performing methylation analysis of nucleic acid molecules derived from the plurality of modified DNA fragments.

A method of mitigating and rehabilitating soil and land areas contaminated with forever chemicals such as PFAS (perfluoroalkyl and polyfluoroalkyl) based toxins involves manipulating genetic nucleotides of carrier organisms for expressing an enzyme reactive with the forever chemicals for rendering them benign. The carrier organisms pass the modified DNA that enables the enzyme to subsequent generations for maintaining an active population of enzyme-expressing organisms, typically worms, once introduced.

The application discloses a kind of lysosome-targeting protein modulators based on upconversion nanomaterial-DNA nanostructure and preparation method thereof.The lysosome-targeting protein modulators are combined by covalent bond from carboxyl functionalized upconversion nanoparticles and amino-modified DNA nanostructure, mainly enter cancer cells through folate receptor, after near infrared light illumination and glutathione response, the target protein combined is transported into lysosome and degraded.The lysosome-targeting protein modulators of the application have the advantages of light affinity, high controllability and storage stability, are easy to be metabolized by organism, and have good application prospect in the field of cell biology and cancer targeted therapy.

Provided herein are methods of analyzing DNA comprising degrading forms of DNA sequences that are prevalent in healthy subjects; and detecting sequences that are not degraded. Some such methods facilitate detection of aberrant forms of DNA.

Provided herein are epigenetic-modifying DNA-targeting systems, such as CRISPR-Cas / guide RNA (gRNA) systems, for the transcriptional repression of Hepatitis B viral (HBV) genes to promote a cellular phenotype that leads to the reduction of HBV infection. In some embodiments, the epigenetic-modifying DNA-targeting systems bind to or target a target site of at least one gene or regulatory element thereof in a Hepatitis B viral DNA sequence in cell. In some aspects, the provided systems relate to the transcriptional repression of one or more Hepatitis B viral gene and / or regulatory element thereof. In some aspects, also provided herein are methods and uses related to the provided compositions, for example in repressing Hepatitis B viral replication and expression in connection with Hepatitis B infections.

The present invention relates to a surface-modified DNA-based nano metal-organic framework (Metal-Organic Framework, abbreviated as MOF) and its application. The organic framework serves as a universal platform for activatable photodynamic cancer therapy, that is, a DNA-functionalized nano metal-organic framework serves as a universal platform for activatable photodynamic cancer therapy. Specifically, it uses an iron-based MOF to load different types of photosensitizers, and utilizes the Fenton-like reaction participated by ferric iron in the tumor microenvironment to specifically release and activate the photosensitizers at the tumor site, improving the therapeutic effect. The surface-modified DNA can not only improve the biocompatibility of the nanoparticles but also effectively prevent drug leakage during blood circulation, thereby reducing the impact on normal tissue cells. It is a targeted therapeutic nano drug and belongs to the technical fields of material chemistry and nano medicine.

An epigenetically modified DNA targeting system, such as a CRISPR-Cas / guide RNA (gRNA) system, is provided that binds to or targets a target site in a gene or a regulatory element thereof in a T cell. In some aspects, the provided epigenetically modified DNA targeting system modulates T cell function, such as T cell phenotype or activity. In some aspects, also provided herein are methods and uses related to the provided compositions, such as modulating T cells, including in combination with methods of adoptive T cell therapy.