39 results about "Nucleic acid hybridisation" patented technology

The present disclosure provides systems and methods for transposing a cargo nucleotide sequence into a target nucleic acid site in a target nucleic acid. These systems and methods may comprise a double-stranded nucleic acid comprising the cargo nucleotide sequence, wherein the cargo nucleotide sequence interacts with a transposase recognition complex, an effector complex comprising an effector and at least one engineered guide polynucleotide that hybridizes to the target nucleic acid, and the transposase recognition complex wherein the transposase recognition complex recruits the cargo nucleotide to the target nucleic acid site.

Multivalent binding compositions including a particle-nucleotide conjugate having a plurality of copies of a nucleotide attached to the particle are described. The multivalent binding compositions allow one to localize detectable signals to active regions of biochemical interaction, e.g., sites of protein-protein interaction, protein-nucleic acid interaction, nucleic acid hybridization, or enzymatic reaction, and can be used to identify sites of base incorporation in elongating nucleic acid chains during polymerase reactions and to provide improved base discrimination for sequencing and array based applications.

Multivalent assemblies for target hybridization are described. The multivalent assemblies include oligonucleotide sets that hybridize to a target nucleic acid to permit capture of the target nucleic acid. In an embodiment, the multivalent assemblies are heteromultivalent such that the oligonucleotide sets include different oligonucleotides that bind to different regions of the target nucleic acid.

A CRISPR-Cas system and applications thereof are provided, also related to a Cas protein, a fusion protein and a guide polynucleotide. The Cas protein has at least 50% sequence identity compared to the sequence set forth in any one of SEQ ID NOs: 1-184, 426-428. The fusion protein comprises the Cas protein fused to a protein domain and / or a polypeptide tag. The guide polynucleotide comprises a direct repeat sequence having at least 70% sequence identity to any one of SEQ ID NOs: 185-199, 245-247, 287-303, 369 and 370-373, and a guide sequence engineered to hybridize to a target nucleic acid. The CRISPR-Cas system comprises a Cas protein having at least 90% sequence identity compared to the sequence set forth in any one of SEQ ID NOs: 1-184, 426-428, or a nucleic acid encoding thereof, and a guide polynucleotide or a nucleic acid encoding thereof.

The present invention relates to a method for preparing an in vitro evolution-basedhotspot-derived peptide-nucleic acid hybrid molecule. According to the method of the present invention, a hotspot-derived peptide-nucleic acid hybrid molecule that can bind with high affinity to viruses and effectively block the binding between the virus and the receptor can be rapidly prepared and screened in response to various virus mutations among numerous candidates. Therefore, it can be used very effectively for the development of therapeutics against various viral mutations.

The invention provides a method for detecting escherichia coli O157 based on a CRISPR (clustered regularly interspaced short palindromic repeats) technology. The method comprises the following steps: S1, amplifying a sample to be detected to obtain an amplification product; amplification of the to-be-detected sample is to amplify the to-be-detected sample by using the RPA amplification primer group; s2, contacting the amplification product with Cas protein, sgRNA and a single-stranded nucleic acid detector, and detecting a detectable signal generated by cutting the single-stranded nucleic acid detector by the Cas protein, so as to detect the escherichia coli O157, the sequences of the RPA amplification primer group are as shown in SEQ ID No.2 and SEQ ID No.7, and the sequence of a target region hybridized with target nucleic acid in the sgRNA is as shown in SEQ ID No.9. The method disclosed by the invention has the advantages of good inclusiveness, strong specificity, high sensitivity and the like, and has a wide application prospect.

Provided are synthetic strands for nucleic acid sequencing. In some embodiments, the strands include a plurality of rotatable molecular disks. The plurality of rotatable molecular disks comprises molecular disks each comprising a first moiety that binds to adenine (A), a second moiety that binds to cytosine (C), a third moiety that binds to guanine (G), and a fourth moiety that binds to thymine (T), uracil (U), or both (T / U). The first, second, third, and fourth moieties are spaced about the perimeter of the molecular disk. The molecular disks enable hybridization of the synthetic strand to a nucleic acid, where the rotational positions of the molecular disks indicate the sequence of the nucleic acid. Also provided are methods of using the synthetic strands, as well as related compositions, kits, and nucleic acid sequencing systems. Stem-loop structure-based sequencing methods and related compositions, kits, and nucleic acid sequencing systems are also provided.

The present invention provides oligomeric compounds. Certain such oligomeric compounds are useful for hybridizing to a complementary nucleic acid, including but not limited, to nucleic acids in a cell. In certain embodiments, hybridization results in modulation of the amount activity or expression of the target nucleic acid in a cell.

The application provides a method for detecting Staphylococcus aureus based on CRISPR technology, and the method comprises the following steps: S1, amplifying a to-be-detected sample to obtain an amplification product; the to-be-detected sample is amplified by using an RPA amplification primer group; S2, contacting the amplification product with Cas protein, sgRNA and single-stranded nucleic acid detector, detecting a detectable signal generated by the Cas protein cutting the single-stranded nucleic acid detector, and thus detecting Staphylococcus aureus; the sequence of the RPA amplification primer group is shown as SEQ ID No. 3 and SEQ ID No. 7, and the sequence of the target region in the sgRNA for hybridizing with the target nucleic acid is shown as SEQ ID No. 9. The method has the advantages of good inclusivity, strong specificity, high sensitivity and the like, and has a wide application prospect.

Provided herein are methods, compositions, and kits for the spatial analysis of target nucleic acids in biological samples by their 5′ end. An exemplary method includes: contacting a biological sample with a primer that hybridizes the target nucleic acid; hybridizing the primer to the target nucleic acid and extending using the target nucleic acid as a template to generate an extension product; incorporating a non-templated polynucleotide sequence including at least three nucleotides to the 3′ end of the extension product; hybridizing the polynucleotide sequence of the extension product to a capture domain on an array including a plurality of capture probes that include a spatial barcode and the capture domain; and determining the spatial barcode sequence, or complement thereof, and all or a portion of the extension product, or complement thereof, and using the determined sequences to determine the location of the target nucleic acid in the biological sample. The non-templated polynucleotide sequence may be C-C-C or C-G-C introduced by terminal transferase or may be introduced by SMART technology using reverse transcriptase.

The invention provides a method for detecting staphylococcus aureus based on a CRISPR (clustered regularly interspaced short palindromic repeats) technology. The method comprises the following steps: S1, amplifying a sample to be detected to obtain an amplification product; amplification of the to-be-detected sample is to amplify the to-be-detected sample by using the RPA amplification primer group; s2, enabling the amplification product to be in contact with Cas protein, sgRNA and a single-stranded nucleic acid detector, and detecting a detectable signal generated by cutting the single-stranded nucleic acid detector by the Cas protein, so as to detect the staphylococcus aureus, the sequences of the RPA amplification primer group are as shown in SEQ ID No.3 and SEQ ID No.7, and the sequence of a target region hybridized with target nucleic acid in the sgRNA is as shown in SEQ ID No.9. The method disclosed by the invention has the advantages of good inclusiveness, strong specificity, high sensitivity and the like, and has a wide application prospect.

This invention relates to the field of biosensors and microfluidics, and particularly to a method for treating interfering DNA based on an ultrasonic-driven device. The method includes an ultrasonic-driven device comprising a piezoelectric substrate, Rayleigh wave drive, microfluidic reaction, and drive control module. First, a nucleic acid hybridization system is constructed to achieve initial binding and recognition of interfering DNA. Then, Rayleigh waves are excited to generate physical shearing force. Based on the difference in binding force between the interfering DNA and the target DNA and probe, precise mechanical feature recognition is achieved. By adjusting the radio frequency signal parameters to make the shearing force between the two types of binding forces, the interfering DNA is dissociated and expelled, completing the selective removal process.

The invention discloses a fluorescent microsphere coding method for regulating and controlling FRET efficiency based on nucleic acid and application, and belongs to the technical field of DNA detection. The method comprises the following steps: combining DNA (deoxyribonucleic acid) coding probes with different lengths and two fluorescent probes marked with different fluorescent groups to the surfaces of magnetic beads through nucleic acid hybridization; the FRET efficiency of the two fluorophores is changed, so that the fluorescence intensity change on the carboxyl magnetic beads is realized; the method comprises the following steps: S1, combining a coupling probe with a microsphere: coupling a first coupling probe to the surface of a carboxylated magnetic bead through a carboxyl-amino covalent reaction; s2, hybridization reaction: taking magnetic beads obtained by coupling, adding a DNA coding probe, a first fluorescent probe and a second fluorescent probe, reacting for a period of time in a dark place, and washing to obtain coding microspheres; and S3, decoding: detecting the fluorescence intensity of FAM and Cy3 through a flow cytometry, and decoding according to the position difference of the microspheres in the two-dimensional scatter diagram. According to the invention, coding in two dimensions of wavelength and intensity can be realized.

The present disclosure provides systems and methods for transposing a cargo nucleotide sequence into a target nucleic acid site in a target nucleic acid. These systems and methods can include: a double-stranded nucleic acid comprising the cargo nucleotide sequence, where the cargo nucleotide sequence interacts with a recombinase complex; an effector complex, the effector complex comprising an effector and at least one engineered guide polynucleotide, the at least one engineered guide polynucleotide hybridizing to the target nucleic acid; and the recombinase recognition complex, wherein the recombinase recognition complex recruits the cargo nucleotide to the target nucleic acid site.

The invention relates to the technical field of analysis and detection, in particular to a nucleic acid detection method based on light-excited chemiluminescence. The method is a homogeneous analysis method, target detection can be completed through direct mixing of target nucleic acid and a detection solution, and the method is high in sensitivity, high in detection speed, free of solid biological recognition carriers and complex cleaning and separation steps, easy to operate and low in cost. According to the method, a chemiluminescence technology and a nucleic acid hybridization technology are combined together, the problem of low sensitivity caused by substrate color development is greatly improved, and the method can be universally applied to detection of nucleic acid molecules from different sources and has a wide application prospect.

RNA interference-based nucleic acids encoding double homeobox 4 (DUX4)-encoding guide RNAs (gRNA), or gRNAs that specifically hybridize to a target nucleic acid encoding DUX4 and methods for inhibiting the expression of DUX4 using the gRNAs and Cas13 are provided. The methods have application in the treatment of muscular dystrophies including, but not limited to, facioscapulohumeral muscular dystrophy (FSHD), and cancer, and other disorders associated with elevated DUX4 expression.

The present disclosure relates to a composition for preventing or treating coronavirus infection, including a hotspot-derived peptide-nucleic acid hybrid molecule. It was confirmed that in vitro evolution-based hotspot-derived peptide-nucleic acid hybrid molecule prepared using the method of the present invention has high binding affinity for the RBDs of SARS-CoV-2 VOCs (alpha, beta, gamma, delta, and omicron). In particular, it was found that the greatest binding tolerance was exhibited in the most highly mutated omicron. Furthermore, the hybrid molecule showed high RBD binding affinity in competition with RBD-binding nucleic acid aptamers, macrocyclic peptides, and monoclonal antibodies. The hybrid molecule also exhibited excellent nuclease resistance and serum stability, indicating potential as virus neutralizer in addition to SARS-CoV-2.

Provided is a nucleic acid measurement method for separately measuring a target nucleic acid conjugate in which a target nucleic acid is modified with a conjugate and a target nucleic acid conjugate metabolite which is a part or the whole of the target nucleic acid to which the conjugate is not bonded. The method comprises: a step in which a fluorescently labeled probe that includes a photocrosslinkable artificial nucleic acid is hybridized with the target nucleic acid to form a target nucleic acid-fluorescently labeled probe complex; a step in which the target nucleic acid-fluorescently labeled probe complex is irradiated with ultraviolet ray to form a covalent bond between the target nucleic acid and the fluorescently labeled probe, whereby a target nucleic acid-fluorescently labeled probe conjugate is formed; and a step in which the target nucleic acid-fluorescently labeled probe conjugate is analyzed by high-performance liquid chromatography and a peak corresponding to the target nucleic acid-fluorescently labeled probe conjugate is detected from a fluorescence wavelength emitted by the fluorescently labeled probe.

This invention discloses a low-voltage driven ratiometric bipolar electrode electrochemiluminescence biosensor and its applications. It utilizes an HNCQDs / H2O2 system and Ru(bpy)3... 2+ The / TPrA system serves as a dual-anode signal source, enabling the construction of a novel ratiometric bipolar electrode electrochemiluminescence biosensor for miRNA detection and analysis. By capturing probes that specifically recognize target miRNAs and auxiliary probes modified with ALP-Au NPs, a "Y-shaped" nucleic acid hybridization configuration is formed. Based on enzyme-catalyzed signal amplification strategies and the working principle of bipolar electrodes, quantitative analysis of miRNA-222 can be achieved through signal intensity ratio analysis based on the changes in the intensity of the two ECL signals, reducing the probability of false positives. Furthermore, the selected dual-anode ECL luminescent probes have low excitation potentials, effectively reducing the driving voltage of the ratiometric bipolar electrode electrochemiluminescence biosensor and expanding the selectivity of ECL luminescent probes. This invention offers advantages such as good portability, low cost, and high sensitivity, and has broad application prospects in clinical diagnosis.

A method for identifying a nucleotide in a template nucleic acid by: (a) providing a plurality of primer-template nucleic acid hybrids, wherein the primers have extendable 3' ends; (b) contacting the plurality of primer-template nucleic acid hybrids with: (i) a blocked nucleotide to produce a first subset of primer-template nucleic acid hybrids comprising a blocked nucleotide at the 3' end, and (ii) a ternary complex inhibitor to produce a second subset of primer-template nucleic acid hybrids comprising a ternary complex inhibitor; (c) forming ternary complexes each comprising a polymerase, a primer-template nucleic acid hybrid of the first subset, and a cognate nucleotide; and (d) detecting the ternary complexes, thereby identifying a nucleotide in the template nucleic acid.

The present disclosure provides methods and devices for simultaneous identification of a plurality of target nucleic acid sequences in a single sample chamber that includes an addressable array of nucleic acid probes attached to a solid surface. Addressable signals can be generated and measured, in real-time, upon hybridization of target sequences at the individual probe locations within the array while the temperature of the system is varied. Such generated signals, as a function temperature, can then be used to compute the properties of nucleic acid hybridization at each addressable location which is ultimately utilized to estimate the sequence of the target nucleic acids. In particular, an integrated semiconductor biosensor array device can be used to measure the addressable signals.

Binding complexes including a detectable nucleotide conjugate having a plurality of copies of a nucleotide attached to the particle are described. The binding complexes allow one to localize detectable signals to active regions of biochemical interaction, e.g., sites of protein-protein interaction, protein-nucleic acid interaction, nucleic acid hybridization, or enzymatic reaction, and can be used to identify sites of base incorporation in elongating nucleic acid chains during polymerase reactions and to provide improved base discrimination for sequencing and array-based applications.

A method includes inserting a fluid including a sample into a cartridge, lysing the sample in an extraction chamber to release nucleic acids of interest from the sample, collecting a first set of hybridized magnetic particles hybridized to a first type of nucleic acids of interest by: hybridizing the first type of nucleic acids of interest with an oligonucleotide attached to magnetic particles, holding the first set of hybridized magnetic particles on a side of the extraction chamber, suspending the first set of hybridized magnetic particles with a wash buffer, transporting the first set of hybridized magnetic particles to a detection chamber, collecting a second set of hybridized magnetic particles hybridized to a second type of nucleic acids of interest, amplifying the sets of hybridized magnetic particles, and detecting a plurality of amplification products indicative of the presence, absence, or amount of the types of nucleic acids of interest.

The invention belongs to the technical field of medical detection equipment, and particularly discloses a nucleic acid detection device. The device comprises a pathogen enrichment unit which is configured to enrich pathogens in a sample to be detected and comprises a sample pool and an enrichment part; the nucleic acid extraction unit is communicated with the pathogen enrichment unit, comprises a nucleic acid releasing agent chamber in which a nucleic acid releasing agent is preset, and is configured to release and dissolve the nucleic acid of the pathogen enriched by the enrichment part; the hybridization sample preparation unit is communicated with the nucleic acid extraction unit and is configured to prepare a single-stranded nucleic acid sample to be hybridized; and the hybridization and detection result display unit is communicated with the hybridization sample preparation unit and is configured to hybridize the single-stranded nucleic acid sample to be hybridized and display a detection result. On the basis of relatively high precision and accuracy, the self-inspection of the viruses and bacteria by the user can be realized without depending on laboratory environment and equipment and having operation speciality.

Multivalent assemblies for target hybridization are described. The multivalent assemblies include oligonucleotide sets that hybridize to target nucleic acids, allowing for capture of the target nucleic acids. In one embodiment, the multivalent assemblies are heteromultivalent, such that the oligonucleotide sets include different oligonucleotides that bind to different regions of the target nucleic acid.

The invention discloses an intracellular glycosylated RNA (Ribonucleic Acid) analysis method for modifying a retardation nucleic acid hybridization technology. Analysis detection and in-situ imaging of intracellular glycosylated RNA (glycoRNAs) are realized on the basis of structural transformation of a target nucleic acid mediated fluorescent substrate nucleic acid chain. Specifically, a substrate chain with toehold is designed according to a sequence of microRNA-21 and a glycosylation modification site of the microRNA-21, and the sequence of the substrate chain contains a specific sequence of a Pepper aptamer capable of being combined with a fluorescent small molecule dye HBC. And the sequence design of the substrate meets the condition that when the substrate is hybridized with glyco-microRNA-21 to a glycosylation modification site, the structure of the generated hybridized compound can just generate an 'ON' fluorescence signal. The hybridization of the substrate sequence and the microRNA-21 is full hybridization, and the formed hybridization product cannot generate a fluorescence signal. According to the invention, the detection of low-abundance glycoRNAs under the interference of a large number of non-glycosylated RNAs is realized, and the intracellular in-situ imaging of the glycoRNAs is realized.

The present disclosure relates to a composition for preventing or treating coronavirus infection, including a hotspot-derived peptide-nucleic acid hybrid molecule. It was confirmed that in vitro evolution-based hotspot-derived peptide-nucleic acid hybrid molecule prepared using the method of the present invention has high binding affinity for the RBDs of SARS-COV-2 VOCs (alpha, beta, gamma, delta, and omicron). In particular, it was found that the greatest binding tolerance was exhibited in the most highly mutated omicron. Furthermore, the hybrid molecule showed high RBD binding affinity in competition with RBD-binding nucleic acid aptamers, macrocyclic peptides, and monoclonal antibodies. The hybrid molecule also exhibited excellent nuclease resistance and serum stability, indicating potential as virus neutralizer in addition to SARS-COV-2.

The present invention relates to an HRCA method comprising the following steps: a) obtaining a nucleic acid structure formed by hybridization of at least one padlock with a target nucleic acid; b) circulating the hybridized padlock; c) subjecting the ligated circulated padlock probe to rolling circle amplification (RCA) and, if necessary, performing an MDA reaction using short primers; and d) detecting the target nucleic acid by detecting the amplified product from step c), preferably by a sequence-specific detection method.

Disclosed here is a device and method to detect the presence or absence of a target molecule by detecting fluorescence resonance energy transfer (FRET) signal generated when two labeled probes bind to the target molecule. The method is a sandwich enzyme-linked immunosorbent assay (ELISA) or a nucleic acid hybridization assay.