67 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 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).

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 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.

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.

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.

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.

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 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.

A modified DNA polymerase includes a DNA polymerase fragment and a G-quadruplex binding peptide fused to an N-terminal of the DNA polymerase fragment.

We describe compositions of DNA donor molecules compatible with various gene insertion methods, offering key advantages in delivery, nuclear localization, and immune evasion. The modified DNA donor molecules are predominantly single-stranded but include short regions of double-stranded DNA to reconstruct enzyme binding sites, and can be combined with other methods to enhance donor recruitment to the cell nucleus.

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.

A composition and method for detecting mesenchymal stem cells using dual markers include magnetic beads bound to a CD73 antibody, a CD44 antibody-modified DNA probe with SH- (C6 Spacer) modification at the 5' end, a signal strand modified with NH2- (C6 Spacer) at the 5' end, a primer modified with Biotin- (C6 Spacer) at the 5' end, and streptavidin-modified magnetic beads. Compared with traditional detection and analysis methods, coating the surface of magnetic beads with specific antibodies, streptavidin, and other functional molecules facilitates the enrichment of target molecules in the presence of complex components such as biochemical samples, effectively improving the efficiency and speed of mesenchymal stem cell and signal product enrichment. Simultaneously, it helps to transfer nucleic acid assembly and detection reactions to a simpler solution environment, reducing background signal interference and thus improving the sensitivity and specificity of the analysis and detection.

Provided are epigenetic-modifying DNA-targeting systems, such as CRISPR-Cas / guide RNA (gRNA) 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 modulate a T cell function, such as a T cell phenotype or activity. In some aspects, also provided herein are methods and uses related to the provided compositions, for example in modulating T cells including in connection with methods of adoptive T cell therapy.

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 invention relates to the technical field of analytical chemistry, and particularly discloses a double-signal reverse change ratio type up-conversion nanoprobe for lead ion detection as well as a preparation method and application of the double-signal reverse change ratio type up-conversion nanoprobe. The double-signal reverse change ratio type up-conversion nanoprobe for lead ion detection comprises an up-conversion nanoparticle coated with polyacrylic acid and a DNA (Deoxyribose Nucleic Acid) aptamer modified by Cy3, wherein the Cy3 modified DNA aptamer is used as a specific recognition unit of Pb < 2 + >, the Cy3 is used as an energy receptor, and the polyacrylic acid coated up-conversion nanoparticles are used as an energy donor. According to the double-signal reverse change ratio type up-conversion nanoprobe, the DNA aptamer is induced to generate conformational change through specific binding of the DNA aptamer and Pb < 2 + >, the distance between up-conversion nanoparticles and Cy3 is shortened, and a ratio type detection signal is constructed through quenching of an emission peak of the up-conversion nanoparticles at 540 nm and enhancement of an emission peak of Cy3 at 565 nm; finally, high-sensitivity and high-selectivity detection on trace Pb < 2 + > is realized, and the detection sensitivity is improved.

The invention discloses a primer probe composition for detecting the DNA methylation level of a TMEM213 gene promoter region and application of the primer probe composition, and belongs to the technical field of biology. The primer probe composition is designed aiming at a CpG site of a human TMEM213 gene promoter region, after sample DNA is modified through bisulfite, methylation sites of the gene promoter region in the modified DNA are detected through methylation specific real-time fluorescent quantitative PCR, and accurate quantitative analysis of the methylation level of the TMEM213 gene target CpG site in the sample DNA can be achieved on the basis of a Ct value obtained through amplification. The primer probe composition can be specifically combined with a target methylated DNA sequence, methylated and non-methylated templates can be efficiently distinguished, the detection sensitivity is high, and the repeatability is good. The primer probe composition is combined with a primer probe combination of a reference gene ACTB for use, and can be applied to preparation of a TMEM213 gene methylation level detection kit.

The invention relates to a method for synthesizing spherical nucleic acid by reducing gold trichloride through alkynyl-modified DNA. The method comprises the following steps: preparing a gold trichloride aqueous solution, an alkynyl-modified DNA solution, a poloxamer 188 aqueous solution and an HEPES solution; adding a gold trichloride aqueous solution, an alkynyl modified DNA solution and a poloxamer 188 aqueous solution into an HEPES solution, and reacting to obtain the spherical nucleic acid of the DNA modified gold nanoparticles. According to the method, the alkynyl-modified DNA is used for reducing gold trichloride to synthesize the spherical nucleic acid in one step for the first time, the use of the alkynyl-modified DNA realizes rapid one-step synthesis of the DNA-modified gold nanoparticles, and the synthesis and modification processes of the spherical nucleic acid are greatly simplified. The preparation method is simplified, and the reaction time is shortened.

The application discloses a controllable assembly method of a nano-gold-DNA probe based on protein-assisted freezing acceleration, which is realized by mixing a nano-gold solution with sufficient protein solution and thiol-modified DNA solution, and then performing freezing, thawing and centrifugation steps. The addition of sufficient protein forms a "protein crown" on the surface of the nano-gold, which protects the nano-gold from aggregation during the freezing process by steric hindrance. The thiol-modified DNA can be connected to the surface of the nano-gold through an Au-S bond. The process is greatly accelerated by the "limiting effect" of the ice crystals formed in the freezing process. Meanwhile, the density of the DNA on the surface of the nano-gold can be controlled by adjusting the amount of the added DNA. The assembly method has the advantages of simple operation, short time consumption and adjustable DNA density, and provides a new general strategy for the assembly and control of probes in the field of biochemical detection.

Provided herein are methods for treating chronic hepatitis B (CHB) infections in human subjects. In some embodiments, the methods involve administering a lipid nanoparticle carrying an epigenetic-modifying DNA targeting system composed of polynucleotides for targeting repression of total Hepatitis B Viral (HBV) transcripts. Also provided is a lipid nanoparticle carrying the polynucleotides.