24 results about "Deoxyribozyme" patented technology

The invention relates to a normal temperature amplification-CRISPR Cas12a one-pot method detection system and a nucleic acid detection method based on the detection system. The detection system consists of a normal-temperature nucleic acid amplification system (unit I) which focuses on target signal amplification and a CRISPR Cas12a system (unit II) which focuses on target recognition and signal output. By means of a self-driven and deoxyribozyme-mediated reaction equilibrium transfer strategy, the equilibrium in the initial stage of the reaction tends to the unit I, and the equilibrium in the middle and later stages of the reaction tends to the unit II, so that the compatibility of the two units is remarkably improved. By means of the system, high-sensitivity and high-specificity portable one-pot detection of nucleic acid targets can be achieved.

The application discloses a method for imaging magnesium ions in living cells based on chemically modified deoxyribozyme, and belongs to the technical field of metal ion imaging. The method comprises the following steps: chemically modifying deoxyribozyme 10-23, screening and obtaining deoxyribozyme 10-23 (CaBn) which is modified with carboxyl (Ca) at the 8th position of the catalytic ring and benzene ring (Bn) at the 12th position; performing biochemical characterization and comparison; constructing a fluorescence sensor responding to magnesium ions; characterizing the performance; and applying the fluorescence sensor to imaging magnesium ions in living cells. Compared with the wild type, the application can catalyze the cleavage reaction under the condition of lower concentration of magnesium ions; therefore, the fluorescence sensor constructed from the deoxyribozyme can more effectively respond to lower concentration of magnesium ions in vitro, and the detection limit reaches 0.03 mM; in addition, the fluorescence sensor can perform fluorescence imaging on endogenous magnesium ions in living cells, and provides a new molecular tool for the field of molecular biology.

The application discloses a kind of photoactivation manganese ion bridging type deoxyribozyme probe preparation and its application in Cistanche inhibiting acetylcholinesterase activity.The probe is pSD@MnO2 Composite structure, which is composed of manganese dioxide nanosheet and functional nucleic acid assembly adsorbed on its surface, the assembly includes S chain labeled with fluorescence / quenching group and containing deoxyribozyme cleavage site, and pD chain partially hybridized with S chain and containing photocleavage site.Its preparation includes nucleic acid probe assembly, MnO2 Nanosheet complexing and purification steps.When applied, AChE catalytic product decomposes MnO2, releases Mn 2+ And nucleic acid chain; after centrifugation, pD chain is cut by ultraviolet light activation trigger, and released DNAzyme is cut under the assistance of Mn 2+ It produces amplified fluorescence signal; by comparing signal change before and after adding inhibitor, inhibition activity can be quantitatively evaluated.The application combines photocontrol activation with enzyme cascade amplification, realizes high sensitivity and operation synchronism of detection process, and is especially suitable for high-throughput rapid screening of AChE inhibitor in Cistanche and other natural products.

The invention discloses a glyphosate fluorescence detection method based on RNase H cycle amplified magnetic bead-aptamer-Cas9 / deoxyribozyme, belongs to the technical field of environmental monitoring and biological detection, and can realize high-sensitivity and rapid quantitative detection of glyphosate residues in soil, water bodies and agricultural products. According to the method, the glyphosate is combined with an aptamer to release complementary DNA (cDNA) combined with the aptamer, then signal cascade amplification is realized through RNase H-mediated cDNA cyclic utilization, the anti-interference advantage brought by magnetic bead separation is combined, and the known characteristics of Cas9 endonuclease and DNAzyme in the aspects of nucleic acid recognition and cutting are used as signal amplification and transduction means, so that the detection sensitivity of the glyphosate is improved, and the detection sensitivity of the glyphosate is improved. Therefore, trace detection of glyphosate is realized. According to the invention, sensitive detection of other targets can be realized by replacing nucleic acid aptamers capable of specifically recognizing other targets and correspondingly adjusting the sequence of cDNA to maintain base pairing.

The invention belongs to the technical field of agricultural biological protection, and discloses a targeted screen tube-vector insect lysozyme-nano magnesium compound system and application thereof in prevention and control of areca yellows, the compound system comprises an in-plant treatment module and a propagation blocking module, the in-plant treatment module comprises a screen tube targeted delivery unit, and the propagation blocking module comprises a propagation blocking unit. The sieve tube targeted delivery unit is formed by combining betel nut sieve tube navigation peptide ArePEP1 and a lysozyme-nano magnesium complex through a non-covalent bond; the propagation blocking module comprises an antifeedant micro-capsule activated by salivary enzyme of a vector insect, and phytoplasma deoxyribozyme PdeA and an insect antifeedant are encapsulated in the antifeedant micro-capsule; the target sequence of the phytoplasma deoxyribozyme PdeA is 5 '-GGAAACAG-3'; the in-plant treatment module and the propagation blocking module are jointly loaded on the same delivery vehicle. According to the areca yellows prevention and control compound system and the application method, pathogens in plants can be synchronously inhibited, insect propagation can be blocked, and high-altitude accurate delivery can be realized.

The invention discloses a droplet microfluidic detection system for detecting pathogenic bacteria based on a deoxyribozyme probe, which belongs to the technical field of microbiological detection, and mainly comprises an automatic sample introduction device, a microfluidic droplet generation chip, a capillary tube-laser induced fluorescence detector and a signal acquisition and analysis device, the pathogen detection process comprises the following steps: respectively and automatically injecting an actual water sample, a deoxyribozyme probe, liquid drop oil, a pathogen lysis solution and a reaction buffer solution into the microfluidic liquid drop generation chip by the automatic sample injection device, stably generating liquid drops, and detecting the liquid drops through the capillary tube-laser induced fluorescence detector, and the signal acquisition and analysis device acquires and analyzes the fluorescence signal. The method has the advantages of high detection flux, high stability, high sensitivity, rapidness and the like, and can be applied to the field of environmental microbiological detection.

The application discloses a double-stranded nucleic acid detection composition based on lambda exonuclease and deoxyribozyme and application thereof, and belongs to the technical field of biological technology, and aims to efficiently and sensitively detect double-stranded nucleic acid. The double-stranded nucleic acid detection composition based on lambda exonuclease and deoxyribozyme comprises a double-stranded nucleic acid detection probe, lambda exonuclease and a reporter molecule, the double-stranded nucleic acid detection probe comprises one long single-stranded DNA named recognition chain R and n short single-stranded DNAs named Dz; the recognition chain R comprises a target binding domain and n Dz binding domains arranged in sequence from 5' to 3' in order; the 5' end of the recognition chain R is modified with a functional group; the Dz is a deoxyribozyme capable of cleaving the reporter molecule when the Dz is single-stranded, and the 5' end of the Dz is modified with sulfur. The application has high sensitivity and specificity, and the detection limit can be as low as a femtomole.

This invention belongs to the medical field and provides a calcium carbonate manganese nanoprobe for in situ imaging detection of vascular epidermal growth factor, its preparation method, and its application. In this strategy, a DNA strand (P1 and P2) containing a protein recognition antibody, a protein recognition sequence, and a deoxyribonuclease, along with a molecular beacon DNA strand (H1), is stirred in a DMEM cell culture medium containing calcium chloride and manganese chloride to prepare the calcium carbonate manganese nanoprobe. In acidic tumor cells, this nanoprobe releases P1, P2, H1, and manganese ions. P1 and P2 can simultaneously recognize proteins, forming a complete deoxyribonuclease center, which catalyzes the cleavage of H1 under the action of the cofactor manganese ions, generating a fluorescent signal. This strategy achieves specific in situ dynamic imaging of low-abundance proteins in vivo and has great potential in biomedical research.

The invention discloses deoxyribozyme targeting enolase 1 and application of deoxyribozyme in preparation of a gene medicine for treating rheumatoid arthritis, and belongs to the technical field of biology. On the basis of a mouse ENO1 mRNA secondary structure, 15 deoxyribozyme is designed and screened, Dz8 has the strongest ENO1 mRNA cutting capacity, and the expression level of the gene can be effectively reduced; meanwhile, the Dz8 has Mg-dependent efficient ENO1 mRNA cleavage activity and time-dependent multi-turnover kinetic characteristics, ENO1 expression can be remarkably inhibited at the gene and protein levels, then inflammatory factor secretion is reduced, the glycolysis level is regulated and controlled, and the Dz8 has important application prospects in the field of rheumatoid arthritis gene therapy. The invention provides a new target and a treatment strategy for rheumatoid arthritis gene therapy, and Dz8 is expected to become a potential tool for rheumatoid arthritis gene therapy.

This invention discloses a deoxyribozyme probe for recognizing strontium ions and its applications, belonging to the field of deoxyribozyme probe technology. The nucleotide sequence of the deoxyribozyme probe provided by this invention is any one of the sequences shown in SEQ ID NO:1 to SEQ ID NO:9. Specifically, it is a cis-structured deoxyribozyme probe composed of a deoxyribozyme and a substrate chain with a fluorescent group and a quenching group. The substrate chain sequence is shown in SEQ ID NO:10, and the deoxyribozyme sequence is any one of the sequences shown in SEQ ID NO:17 to SEQ ID NO:25. The deoxyribozyme probe provided by this invention can efficiently and specifically recognize strontium ions, thereby achieving accurate detection of the target ion, with a strontium ion response concentration of 2 mM or higher. This probe has good application prospects in the fields of biosensing and ion detection.

The invention discloses a deoxyribozyme probe capable of specifically distinguishing a G-quadruplex and a single-strand structure of a G-rich sequence and application of the deoxyribozyme probe, and belongs to the technical field of deoxyribozyme probes. The method comprises the following steps: firstly, connecting a phosphorylated library enzyme chain with a substrate chain containing RNA (Ribonucleic Acid) modification sites to construct a linear DNA (Deoxyribose Nucleic Acid) library; then, under the condition that the pH is 7.4, co-incubating the library and a G-rich sequence in a single-chain state, separating and purifying a DNA strip for cutting an RNA site in the library, recovering a positive-sense strand after PCR amplification, and carrying out multiple rounds of screening to obtain the chain deoxyribozyme probe. The obtained probe can specifically detect a G-rich sequence in a single-chain form, has no response to a G-quadruplex structure, and has a good application prospect in the field of biosensing.

The present application provides a method for regulating the activity of functional nucleic acid, which comprises modifying one or more bases of the functional nucleic acid to inactivate the functional nucleic acid. Preferably, the functional nucleic acid or the substrate of the functional nucleic acid is modified to inactivate the functional nucleic acid, and then the modified group is removed to activate the functional nucleic acid. The present application also provides a deoxyribozyme molecular probe, which comprises domain a, domain b and domain c. The deoxyribozyme molecular probe is obtained by screening 6 A closed Dz, which realizes m 6 A high-efficiency and sensitive detection of demethylase and screening and detection of inhibitor small molecule drugs.

The invention relates to the technical field of molecular detection, in particular to deoxyribozyme for detecting BK polyoma virus and application of deoxyribozyme. The deoxyribozyme comprises a catalytic core and a binding arm connected with the 5'end and / or the 3 'end of the catalytic core, wherein the catalytic core has the activity of cutting nucleic acid. The invention further provides a method for detecting the BK polyomavirus, the method comprises the step that a sample to be detected makes contact with the deoxyribozyme, by means of the nucleic acid detection method, the BK polyomavirus as low as 0.1 aM can be detected, and extremely high sensitivity and specificity are achieved.

The present invention is directed to a composition for a target sequence detection in double- stranded DNA, wherein the composition comprises i) a first oligonucleotide and a second oligonucleotide for a multi-component deoxyribozyme (MNAzyme), wherein said first and second oligonucleotides are capable of forming substrate binding arms, a catalytic core, and target-binding arms of said MNAzyme, said target-binding arms being complementary to said target sequence in double-stranded DNA, ii) a peptide nucleic acid (PNA) comprising a sequence identical to said target sequence, wherein said PNA is capable of invading the double-stranded DNA at the site of said target sequence and opening the double-stranded DNA by binding to the sequence complementary to said target sequence in said double- stranded DNA leaving said target sequence single-stranded and thus free for said target- binding arms of said MNAzyme to bind, and iii) a substrate oligonucleotide comprising a sequence complementary to the sequence of said substrate binding arms, wherein said substrate binding arms of said MNAzyme engage said substrate oligonucleotide so that said catalytic core of said MNAzyme is capable of modifying said substrate oligonucleotide when said target-binding arms of said MNAzyme bind or are bound to said target sequence in the presence of a cofactor. The present invention is also directed to a kit comprising said composition and to methods of using said composition.

This application belongs to the field of bioanalytical detection technology, specifically disclosing a detection system and method for live Salmonella based on deoxyribonuclease activation. The detection system includes a Sub probe, a deoxyribonuclease probe, a hairpin probe, hyperbranched dendritic nanomolecules, a crRNA / Cas12a binary complex, a fluorescent probe, and magnetic beads. The hyperbranched dendritic nanomolecules include substrate A, substrate B, and a trigger strand B probe. Substrate A consists of an A-F strand probe, an A-Q strand probe, and a helper strand A probe, while substrate B consists of a B-F strand probe, a B-Q strand probe, and a helper strand B probe. This detection system specifically recognizes ribonuclease H2 released only by the metabolism of live Salmonella and utilizes it to activate the Sub-Dz substrate, ensuring from the source that only live bacteria can trigger the subsequent reaction. This effectively solves the problem of interference from dead bacteria, achieving highly sensitive, highly specific, and rapid detection of live Salmonella.

The present invention concerns the field of modified nucleic acids, and in particular aptamer, ribozyme or deoxyribozyme functional nucleic acids, and methods for selecting (SELEX) such modified nucleic acids. The invention concerns nucleobases, nucleosides and nucleotides modified by a particular chemical group, and the uses thereof, in particular in methods for reverse transcription, amplification and selection of functional nucleic acids, regardless of the chemical group attached to the modified nucleic acids to be selected, provided that said group is cleavable via an SPICC reaction.

The invention discloses an endoplasmic reticulum stress responsive gene regulation and control platform and application thereof. The endoplasmic reticulum stress responsive gene regulation and control platform comprises an IRE1 cuttable hairpin element and a gene regulation and control unit, the gene regulation unit comprises any one of deoxyribozyme, antisense oligonucleotide, mRNA (messenger ribonucleic acid) vaccine, CRISPR (clustered regularly interspaced short palindromic repeats) system or RNAi (ribonucleic acid interference) system. According to the endoplasmic reticulum stress responsive gene regulation and control platform disclosed by the invention, the IRE1 cleavable hairpin element is modularly integrated into the DNAzyme system, so that the cleavage activity of the DNAzyme on target RNA (Ribonucleic Acid) can be inhibited; in tumor cells with high endoplasmic reticulum stress, the IRE1 can restore the catalytic activity of the DNAzyme by damaging a hairpin structure, and the activity of the DNAzyme cannot be activated in normal cells with low endoplasmic reticulum stress, so that gene regulation and control of endoplasmic reticulum stress responsiveness are realized. In addition, the platform can realize on-demand accurate regulation and control of endoplasmic reticulum stress of tumor cells.

The application discloses a nano-label-free nucleic acid detection test strip based on a DNA sponge capture probe. The application uses a three-dimensional biomimetic DNA sponge as a test strip detection line capture interface, and utilizes the porous biomimetic structure to realize the synergistic enhancement effect of anti-coffee ring effect, resident reaction solution, prolonged action time of recognition molecules and capture probes, and multi-valent load to improve the capture efficiency. A lossless chain displacement isothermal amplification is matched, and rich guanine amplification products are continuously generated on the premise of not consuming templates. The products and chlorinated hematin or N-methyl mesoporphyrin IX self-assemble to form a deoxyribozyme complex, and double-mode visual detection is completed through substrate catalytic colorimetric or ultraviolet excitation fluorescence. The application does not use nanoparticles labels in the whole process, is simple and convenient to operate, low in cost, green and environment-friendly, and is suitable for instant detection of various nucleic acid targets, and provides a universal high-performance interface and signal output scheme for paper-based biosensing.

The invention discloses a method for detecting micro RNA (Ribonucleic Acid) by using chemically modified multi-component deoxyribozyme and application of the method, and belongs to the field of molecular diagnosis and biosensing. Heterogeneous nucleic acid modification is introduced into a catalytic core region of a deoxyribozyme molecule, and locked nucleic acid modification is introduced into a target binding arm region; heterogeneous nucleic acid modification is used for enhancing the conformational stability and catalytic rate of the catalytic core, and locked nucleic acid modification is used for improving the recognition affinity and specificity of the target binding arm and the target miRNA. The method has the advantages that the detection reaction is realized under the enzyme-free, isothermal and one-pot conditions, and the detection efficiency and sensitivity are remarkably improved. The catalytic rate is increased by 6.5 times after XNA modification is introduced, the overall catalytic efficiency is increased to 760 times after LNA modification is introduced, and the detection sensitivity is increased to 2 pM. The method is flexible in structural design, simple and convenient to operate, high in specificity and suitable for application scenes such as field detection and liquid biopsy, and has a good industrialization prospect.

The invention discloses a deoxyribozyme probe for recognizing strontium ions and application, and belongs to the technical field of deoxyribozyme probes. The nucleotide sequence of the deoxyribozyme probe provided by the invention is any one of SEQ ID NO: 1-SEQ ID NO: 9, in particular to the cis-structure deoxyribozyme probe consisting of deoxyribozyme and a substrate chain with a fluorophore and a quenching group, the sequence of the substrate chain is as shown in SEQ ID NO: 10, and the sequence of deoxyribozyme is any one of SEQ ID NO: 17-SEQ ID NO: 25. The deoxyribozyme probe provided by the invention can efficiently and specifically recognize strontium ions, so that accurate detection of target ions is realized, and the response concentration of the strontium ions is more than 2 mM. The probe has a good application prospect in the fields of biosensing, ion detection and the like.

The application discloses a specific RNA-m 1 The application discloses a chain deoxyribozyme probe for specifically recognizing RNA-m 1 The application discloses a chain deoxyribozyme probe for specifically recognizing RNA-m 1 The application discloses a chain deoxyribozyme probe for specifically recognizing RNA-m

The invention belongs to the technical field of biological analysis and detection, and particularly discloses a live salmonella detection system and method based on deoxyribozyme activation, the detection system comprises a Sub probe, a deoxyribozyme probe, a hairpin chain probe, a hyperbranched dendritic nano-molecule, a crRNA / Cas12a binary compound, a fluorescent probe and magnetic beads, the hyperbranched dendritic nano-molecule comprises a substrate A, a substrate B and a trigger chain B probe, the substrate A is composed of an A-F chain probe, an A-Q chain probe and an auxiliary chain A probe, and the substrate B is composed of a B-F chain probe, a B-Q chain probe and an auxiliary chain B probe. According to the detection system, ribonuclease H2 released only by metabolism of living salmonella is specifically recognized, and is utilized to activate a Sub-Dz substrate, so that the subsequent reaction can be triggered only by viable bacteria from the source, the problem of dead bacteria interference is effectively solved, and high-sensitivity and high-specificity rapid detection of the viable salmonella is realized.

The invention discloses a deoxyribozyme probe for specifically distinguishing inosine and adenosine and application, and belongs to the technical field of deoxyribozyme probes. A substrate chain containing an unmodified RNA site or a substrate chain with an RNA modification site is connected with a DNA library to construct a chain DNA complex, in-vitro screening is carried out, chain deoxyribozyme capable of specifically recognizing an inosine site is obtained through screening, and the catalytic activity of the chain deoxyribozyme is the highest in a publically reported specific recognition RNA modified deoxyribozyme probe. The probe obtained by the invention can specifically detect and recognize sites containing inosine, and has the potential of being applied to the aspect of biosensing.

The invention discloses a multifunctional DNA nano assembly as well as a preparation method and application thereof. The multifunctional DNA nano assembly comprises DNA nano flowers, chemotherapeutic drugs are loaded on the DNA nano flowers, the surfaces of the DNA nano flowers loaded with the chemotherapeutic drugs are coated with metal polyphenol networks, and hyaluronic acid is connected to the surfaces of the metal polyphenol networks; the DNA nanoflower is obtained through a rolling circle amplification reaction, and a repetitive unit sequence of the DNA nanoflower is as shown in SEQ ID NO.3. The preparation method comprises the following steps: firstly, preparing DNA nanoflowers encoding deoxyribozyme sequences through a rolling circle amplification reaction, then loading a chemotherapeutic drug on the DNA nanoflowers, then coating the surface of the DNA nanoflowers with a metal polyphenol network, and finally carrying out hyaluronic acid functionalization, so that the prepared multifunctional DNA nano assembly can enter tumor cells in a targeting manner through hyaluronic acid, and the tumor cells can be targeted through the hyaluronic acid. The combined treatment of gene therapy, chemotherapy and photothermal therapy can be realized under the irradiation of 808 nm laser.