40 results about "Modified dna" patented technology

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 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.

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.

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 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.

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.

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.

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.

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.

The present invention relates to a composition comprising a modified DNA identifier sequence (MoDIS) comprising (i) a verification code nucleotide sequence (VC), (ii) a random index code nucleotide sequence (RIC) and (iii) an annealing site nucleotide sequence (AS) for specific binding to a primer for PCR amplification, wherein (a) AS is linked to an affinity tag or label and / or affinity tag compound for identifying, enriching and / or purifying MoDIS-bound nucleotide sequences; and (b) the nucleotide at the 5 '-terminus of the MoDIS comprises a 5'-azido-modified moiety or a 5 '-alkynyl-modified moiety.

The present invention relates to a method for capturing nucleic acid sequences and maintaining / preserving their epigenetic information, preferably for capturing specific regions of the genome, thereby enriching these regions, while maintaining their epigenetic information with the same probability / efficiency, regardless of whether one or more nucleotides are epigenetically modified. Specifically, the present invention relates to a method comprising the steps of: (i) providing a plurality of nucleic acid molecules, wherein each nucleic acid molecule comprises two regions: a first region and a second region, wherein the base types in the first region and the second region independently provide information about the base types in corresponding loci of one or more original nucleic acid molecules, wherein the first region and the second region in each molecule provided in (i) contain at least one nucleotide at a position corresponding to the same locus in a target region, wherein the locus in the target region is occupied by a modifiable nucleotide; wherein at least two of the plurality of nucleic acid molecules may have different nucleotides at at least one position in the first region corresponding to the same locus in the target region; wherein the locus in the target region is occupied by a modifiable nucleotide, and wherein, in the at least two nucleic acid molecules, the position corresponding to the same locus in the target region in the second region is occupied by the same nucleotide; and (ii) capturing the molecule provided in (i) using at least one capture probe, the capture probe binding to at least a portion of the second region, at least a portion of the second region containing at least one nucleotide located at a position corresponding to a locus occupied by a modifiable nucleotide in the ROI.

The invention provides a TiO2 / ZnS / Au-C3N4 / aptamer PEC biosensor as well as a preparation method and application of the TiO2 / ZnS / Au-C3N4 / aptamer PEC biosensor. Comprising a TiO2 / FTO substrate layer, a ZnS middle layer, an Au-C3N4 selection layer and a DNA aptamer for identifying Abeta42 from bottom to top in sequence. Energy level matching among the three materials is utilized to form the stepped heterojunction, ZnS serves as a middle bridge, separation and enrichment of electrons and holes are promoted, and therefore the photocurrent efficiency is remarkably improved. The sensor is functionalized by a sulfydryl-modified DNA aptamer, and the aptamer is covalently bonded with Au NPs on Au-C3N4 to form an Abeta42 high-specificity selection layer, so that the stability of the sensor is improved, and the number of adsorbable sites is increased; and in consideration of selective detection interference of a complex plasma matrix on A beta42 and a negative signal caused by the selective detection interference of the complex plasma matrix on A beta42, the surface is coated with a layer of mercaptohexanol (MCH), so that the problem of electrode pollution is effectively relieved. The sensor prepared by the invention realizes rapid and accurate detection of the level of Abeta42 in plasma at a cost which is obviously lower than that of the current gold standard.

C-5 propyl-G2-tagged pyrimidine and C-8 propyl-G2-tagged purine nucleoside. These nucleosides are useful as precursors for modified DNA and / or RNA strands. The resulting strands may have non-canonical confirmations (NCCs).

The application discloses a transcytosis DNA nanoframework material, a preparation method and application thereof, and the transcytosis DNA nanoframework material is assembled through a hybridization chain reaction of a DNA-containing polymer as a skeleton and DNA hairpins; the skeleton is prepared from at least acrylamide-modified DNA, diethylaminoethyl methacrylate and 3-methacrylamidophenylboronic acid as raw materials through polymerization and oxidation. The transcytosis DNA nanoframework material integrates transcytosis function and gene silencing function, and can be used for precise delivery and treatment of pancreatic cancer and other solid tumors rich in dense matrix barriers.

This invention discloses a PSA aptamer sensor based on a screen-printed electrode, its preparation, and detection method. In this invention, a composite material of AuNPs and graphene aerogel is used to modify the surface of a screen-printed electrode. The aptamer is modified onto the sensor by forming gold-sulfur (Au-S) bonds with a thiol-modified DNA single-stranded aptamer. Excess gold active sites are blocked with MCH to construct the PSA aptamer sensor. A certain concentration of PSA and UA is selected and incubated with the sensor. The EIS test method is used to detect PSA by recording its impedance changes, and the detection limit of the sensor is obtained. Utilizing the advantages of large-scale mass production and low cost of screen printing technology, combined with the excellent physical and electrical properties of nanomaterials, an electrochemical sensor for the detection of prostate-specific antigen (PSA) is prepared, enabling convenient, efficient, and low-cost detection, providing a new approach for prostate cancer patients to be detected outside of hospitals and laboratories.

The invention discloses a preparation method and application of a novel azobenzene modified nanopore probe, and belongs to the technical field of biological analysis and detection. The preparation method comprises the following steps: dissolving DNAs modified by dibenzocyclooctyne in water to obtain a DBCO-DNA solution; adding a buffer solution into the DBCO-DNA solution, then adding an azide-polyethylene glycol (5)-N-hydroxysuccinimide ester aqueous solution to carry out a click chemical reaction, and purifying to obtain an intermediate product; and adding a 4-aminoazobenzene hydrochloride acetonitrile solution into the intermediate product, and carrying out a coupling reaction under an alkaline condition to obtain the product. The invention also discloses a novel azobenzene modified nanopore probe and application thereof. The novel azobenzene modified nanopore probe can generate nanopore characteristic signals with characteristics and distinguishability, real-time, label-free, damage-free and high-sensitivity detection on enzymes and small molecules can be achieved by combining the azobenzene modified nanopore probe with a nanopore single-molecule detection platform, and the application potential is large.

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 discloses a preparation method of a CTC capture detection kit based on a prostatic cancer specific marker, and relates to the technical field of circulating tumor cell detection. Screening to obtain a membrane protein marker combination which is stably co-expressed in circulating tumor cells and is not expressed in normal peripheral blood nucleated cells, wherein the membrane protein marker combination comprises at least two of PSMA, STEAP1 and NKX3-1; 2, aiming at each member in the membrane protein marker combination obtained in the step 1, screening through an SELEX (systematic evolution of ligands by exponential enrichment) technology to obtain a corresponding DNA (deoxyribonucleic acid) aptamer, and carrying out 5 '-terminal amino modification on the obtained DNA aptamer; 3, providing a solid-phase chip with a micropore array structure, and carrying out polyethylene glycol surface passivation treatment on the inner walls of micropores; and 4, mixing the amino-modified DNA aptamers obtained in the step 2, and fixing the amino-modified DNA aptamers on the inner walls of the passivated micropores obtained in the step 3 in a covalent coupling manner.

The application discloses a primer probe composition for detecting a DNA methylation level in a TMEM213 gene promoter region and application thereof, and belongs to the technical field of biology. The primer probe composition is designed according to a CpG site in a human TMEM213 gene promoter region, and after sample DNA is modified by a bisulfite reagent, methylation-specific real-time fluorescent quantitative PCR is used to detect the methylation site in the gene promoter region of the modified DNA; and based on the Ct value obtained through amplification, the methylation level of the target CpG site of the TMEM213 gene in the sample DNA can be accurately quantitatively analyzed. The primer probe composition can specifically combine with the target methylation DNA sequence, can efficiently distinguish the methylation template from the non-methylation template, has high detection sensitivity and good repeatability. The primer probe composition is combined with a primer probe combination of an internal reference gene ACTB, and can be applied to the preparation of a TMEM213 gene methylation level detection kit.

Disclosed herein is an antibody to PcrV as well as nucleic acid molecules encoding the anti- PcrV antibody and compositins comprising the antibody or nucleic acid molecules encoding the antibody. The disclosure also provides a method of preventing and / or treating a bacterial infection in a subject using the antibody or nucleic acid molecules.