1130 results about "Double strand" patented technology

The invention discloses a method for detecting the concentration of cyclosporine in blood through fluorescence and electrochemical double signals, and belongs to the technical field of biochemical analysis and medical detection. Aiming at the technical problems of insufficient sensitivity and high false positive rate of traditional single-signal detection of blood concentration of cyclosporine, the method comprises the following steps: activating carboxyl microspheres by a coupling agent, fixing double-stranded DNA (deoxyribonucleic acid) formed by a cyclosporine aptamer and an initiator chain, inducing the initiator to release by using a cyclosporine sample, realizing strand displacement through an A2-PEI composite system, and detecting the blood concentration of cyclosporine by using an A < 2 >-PEI composite system. DNA modified cadmium telluride quantum dots are respectively adopted to detect fluorescence signals, eATRP electrochemical signal amplification is carried out to detect electrochemical signals, concentration values obtained by the two signals are verified, and an average value is taken as a result. The method can accurately capture low-concentration cyclosporine signals in blood, eliminates interference of a single method, is mainly used for accurately monitoring the blood concentration of cyclosporine, and provides a reliable detection means for medication safety and curative effect control of cyclosporine in clinical scenes such as organ transplantation.

The disclosure provides, for example, double stranded DNA (dsDNA) molecules comprising one or more chemically modified nucleobases. In some embodiments, the dsDNA molecule is circular and comprises a first strand and a second strand, wherein the first strand comprises one or more chemically modified nucleobases, and the second strand is free of chemically modified nucleobases. In some embodiments, the dsDNA molecule comprises a promoter sequence and an effector sequence that encodes an effector.

The present disclosure relates to isolated nucleic acid molecules comprising at least two double-stranded inhibitory ribonucleic acid (RNA) molecules adapted to silence the same or different genes by RNA interference to enhance silence, thereby modulating gene expression.

The disclosure relates to double-stranded ribonucleic acid (dsRNA) targeting an INHBE gene, and methods of using the dsRNA to inhibit expression of INHBE.

The present disclosure provides compositions comprising nucleic acid partially double- stranded splint adaptors, and methods for preparing nucleic acid libraries using the partially double-stranded splint adaptors. The partially double-stranded splint adaptors can be ligated to double stranded nucleic acid fragments to form library-splint complexes having double stranded nicks, where the nicks can be ligated to form covalently closed circular molecules, which can then be subjected to downstream amplification and sequencing workflows.

The invention belongs to the technical field of microorganisms, and particularly relates to recombinant escherichia coli containing a PDRN sequence and a preparation method thereof, and the preparation method comprises the following steps: S1, preparing a PDRN solution; s2, complementing the double strands of the PDRN into blunt ends; s3, constructing a recombinant plasmid; s4, carrying out mixed culture on a product obtained in the step S3 and escherichia coli DH5alpha; s5, screening the recombinants; s6, carrying out PCR (Polymerase Chain Reaction) amplification, and recording a strain from a fragment with the length of more than 200bp; s7, preparing a seed solution by adopting the strain prepared in the step S6; s8, strain fermentation; and S9, centrifugally collecting thalli. The method comprises the following steps: constructing a recombinant plasmid containing a PDRN fragment, introducing the recombinant plasmid into Escherichia coli, subsequently culturing the recombinant Escherichia coli to copy DNA in the Escherichia coli, extracting a purified exogenous gene fragment to obtain a strain containing a single fragment PDRN sequence, and getting rid of dependence on raw materials during subsequent extraction.

The present invention relates to modulators that can regulate, for example, inhibit the expression and / or activity of an inhibin subunit βE (INHBE), such as double-stranded RNA (dsRNA) active agents or antisense polynucleotide formulations. The present invention also relates to a method for inhibiting the expression and / or activity of INHBE by using such modulators, and a method for preventing and treating INHBE-related diseases in subjects (such as metabolic disorders or obesity or cardiovascular diseases, such as metabolic syndrome).

The invention discloses siRNA for inhibiting GPR75 gene expression and application of the siRNA. The siRNA comprises a positive-sense strand and an antisense strand, and the antisense strand has a nucleotide sequence which is different from a nucleotide sequence shown in any one of SEQ ID NO.109-216 and SEQ ID NO.226-231 by not more than 3 nucleotides; the antisense strand and the positive-sense strand are at least partially complementary to form a double-strand region. The invention provides siRNA for inhibiting GPR75 gene expression, the siRNA is modified to ensure the stability and inhibitory activity of the siRNA, and experiments prove that multiple groups of siRNA have obvious inhibitory activity on GPR75 gene expression, and show application prospects in preparation of drugs for treating or preventing GPR75 expression related diseases in human bodies.

The disclosure features systems and methods for correcting a mutation in the human beta-globin (HBB) gene in a cell or population of cells. The disclosure also features methods of increasing repair of a DNA double stranded break (DSB) in an HBB gene by the homology-directed repair (HDR) pathway. The disclosure also features compositions for use in the methods.

The disclosure provides methods for processing nucleic acid populations containing different forms (e.g., RNA and DNA, single-stranded or double-stranded) and / or extents of modification (e.g., cytosine methylation, association with proteins). These methods accommodate multiple forms and / or modifications of nucleic acid in a sample, such that sequence information can be obtained for multiple forms. The methods also preserve the identity of multiple forms or modified states through processing and analysis, such that analysis of sequence can be combined with epigenetic analysis.

96 New PCT-Patent Application based on EP 24 186 635.9 Insempra GmbH Vossius Ref.: AG4141 PCT S3 Abstract The present invention relates to a linear double-stranded deoxyribonucleic acid (dsDNA) molecule comprising one or more Tus protein (Tus) binding site(s) at the 5'-terminus, one or more Lac repressor protein (LacI) binding site(s) at the 3'-terminus, and a segment comprising a DNA sequence of interest (DOI) between said Tus binding site(s) and said LacI binding site(s). The present invention further relates to a non-naturally or naturally occurring RNA molecule as encoded by or transcribable / transcribed from the linear dsDNA molecule of the invention, and to an expression DNA cassette comprising a promoter, an RBS, a GOI encoding a POI, and one or more LacI binding site(s), wherein said expression cassette does not comprise a terminator sequence between the 3´ end of said GOI and said one or more LacI binding site(s). The present invention also relates to a non-naturally or naturally occurring RNA molecule as encoded by or transcribable / transcribed from said expression cassette. The present invention further relates to method of protecting a linear deoxyribonucleic acid (DNA) molecule from exonuclease degradation by adding one or more Tus binding site(s) at the first terminus of the DNA molecule and adding one or more LacI binding site(s) at the other terminus of the DNA molecule. The present invention further relates to a method of synthesizing a protein of interest (POI) in a cell- free protein synthesis (CFPS) reaction mixture by using the (ds)DNA and / or RNA molecules of the invention, and also to a cell-free biological system or CFPS reaction mixture comprising the (ds)DNA, expression cassette and / or RNA molecules of the invention.

The invention provides double-stranded ribonucleic acid for reducing blood coagulation factor XI expression and a modifier and application of the double-stranded ribonucleic acid. Cell experiment results show that the oligonucleotide duplex can significantly inhibit the expression of the FXI gene, and can be used for developing anticoagulant drugs.

The present disclosure provides double stranded oligonucleotides, compositions, and methods relating thereto. The present disclosure encompasses the recognition that structural elements of double stranded oligonucleotides, such as base sequence, chemical modifications (e.g., modifications of sugar, base, and / or internucleotidic linkages) or patterns thereof, and / or stereochemistry (e.g., stereochemistry of backbone chiral centers (chiral internucleotidic linkages), and / or patterns thereof, can have significant impact on oligonucleotide properties and activities, e.g., RNA interference (RNAi) activity, Ago2 loading, thermal stability, in vivo stability, delivery to tissues and into cells, etc. The present disclosure also provides methods for treatment of diseases, e.g., hepatic diseases, central nervous system (CNS) diseases, etc., using provided double stranded oligonucleotide compositions, for example, in RNA interference.

The invention discloses light-driven toxin-enriched composite hydrogel as well as a preparation method and application thereof. The composite hydrogel comprises light-driven hydrogel and detection hydrogel, the light-driven hydrogel is agarose hydrogel doped with Au (at) Ag core-shell nano particles; the detection hydrogel is a double strand formed by an okadaic acid aptamer and a complementary sequence cDNA thereof, and a double strand formed by a domoic acid aptamer and a complementary sequence DNAzyme thereof, the hairpin H1 is used for modifying a quenching group BHQ2 at the 3'end, the hairpin H2 is used for modifying a fluorophore Cy3 at the neck part, the hairpin H3 is used for modifying a fluorophore FAM and a quenching group BHQ1 at the two ends respectively, and the metal ion doped agarose hydrogel is used for catalyzing DNAzyme cyclic shearing; and the photo-thermal driving unit is positioned on the detection function unit. According to the present invention, with the composite system of upper layer photo-thermal enrichment and lower layer dual-signal detection, the rapid and high-sensitivity simultaneous detection of the okadaic acid and the domoic acid is achieved through the combination of the photo-thermal transpiration effect and the HCR and DNAzyme cyclic shearing reaction;

The invention belongs to the technical field of biological medicines, and particularly discloses a bionic nano-vesicle for regulating and controlling neutrophil phenotype in combination with ultrasound, and a preparation method and application of the bionic nano-vesicle. The bionic nano-vesicle provided by the invention can be broken under the action of high-concentration glutathione existing in a tumor tissue microenvironment to release the sound-sensitive agent and functional siRNA, and under the action of US irradiation, the sound-sensitive agent is activated to generate a large amount of ROS, promote tumor cell DNA breakage, generate a large amount of double-stranded DNA, activate APC and promote IFN-beta release, so that the tumor tissue microenvironment is inhibited. The released functional siRNA can promote the N2-type neutrophile granulocyte to become N1-type neutrophile granulocyte, and the released functional siRNA can prevent the N1-type neutrophile granulocyte from being reversed to the N2-type neutrophile granulocyte again, so that the regulation and control on the phenotype of the neutrophile granulocyte are realized; under catalysis of CAT rich in the thylakoid membrane, H2O2 overexpressed by tumor tissue is decomposed into oxygen, the tumor hypoxia microenvironment can be relieved, and the tumor treatment effect can be improved.

Described herein are methods and compositions for detecting an antigen using at least two antigen binders each comprising partially double-stranded nucleic acids wherein the partially double-stranded nucleic acids can template production of a linear or circular nucleic acid product.

The invention provides a quantitative detection system combining a CRISPR-Cas system with a MetaSPR optical sensor, a detection method and application, and relates to the technical field of biosensing and optical detection. The system comprises a MetaSPR optical sensor, the surface of a gold substrate of the MetaSPR optical sensor is modified with single-stranded DNA H1, the 5'end of H1 is modified with sulfydryl, and H1 is fixed to the gold substrate of the MetaSPR optical sensor through an Au-S bond; h1 is combined with single-chain DNA H2 with gold balls through complementary base pairing to form a double-chain structure, the 5'end of H2 is also modified with sulfydryl, and the gold balls are fixed on the surface of a gold substrate; the CRISPR-Cas reaction system comprises a molecular solution to be detected, Cas protein, a buffer solution, guide RNA (Ribonucleic Acid), nuclease-free water and target DNA (Deoxyribose Nucleic Acid); the CRISPR-Cas reaction system is used for recognizing a target molecule, and when the target molecule is recognized, H1 is cut.

The invention relates to pharmaceutical compositions comprising an iRNA agent, e.g., double stranded ribonucleic acid (dsRNA) agent and methods of using such compositions to treat a bleeding event in a subject having a hemophilia (e.g., with or without inhibitors).

Described herein are methods, systems, and apparatuses for determining a partial consensus sequence of a double-stranded nucleic acid molecule. Both strands of the nucleic acid molecule may be sequenced to generate a sequence of base calls. The base calls may be used to identify sets of concordant and discordant positions. A partial consensus sequence may be generated by using concordant values derived from the concordant positions and discordant values derived from the discordant positions. Also described herein are methods, systems, and apparatuses for determining a consensus sequence of a double-stranded nucleic acid molecule. Both strands of the nucleic acid molecule may be sequenced to generate base calls and quality scores corresponding to each strand. Concordant and discordant positions may be identified using the sequences of base calls. Discordant positions may also use the quality scores and weights. A consensus sequence may be determined using the concordant and discordant positions.

The invention discloses a divalent nucleic acid aptamer fluorescence detection method based on an exonuclease I auxiliary target circulation strategy, and belongs to the field of analysis and detection.The divalent nucleic acid aptamer is a repetitive sequence composed of two monovalent nucleic acid aptamers and can form double-stranded DNA through base complementary pairing with complementary strand cDNA of the divalent nucleic acid aptamer, and the divalent nucleic acid aptamer can be used for fluorescence detection of the divalent nucleic acid aptamer. The method comprises the following steps: taking a bivalent nucleic acid aptamer as a template, forming a DNA silver nanocluster (DNA-AgNCs) under the action of silver nitrate and sodium borohydride, and when the target okadaic acid exists, combining the bivalent nucleic acid aptamer with the target, so that cDNA can be replaced from double-stranded DNA by the target. Exonuclease I is added to perform enzyme digestion on the cDNA and the divalent nucleic acid aptamer, so that the fluorescence intensity of the DNA-AgNCs can be rapidly reduced. The content of the okadaic acid can be judged by detecting the change of the fluorescence intensity. The invention provides a simple, convenient, rapid, high-sensitivity and high-specificity detection method for okadaic acid detection based on a divalent nucleic acid aptamer for the first time.

The disclosure provides, for example, a DNA molecule comprising a first strand of DNA, wherein the first strand is circular and single stranded over at least 90% of its length, and a second strand of DNA covalently linked to the first strand as described herein. The DNA molecule may comprise an effector sequence encoding a therapeutic effector. In some embodiments, the DNA molecule shows improved conversion to double-stranded DNA relative to a DNA molecule that lacks the second strand.

The invention relates to dsRNA capable of simultaneously preventing and treating various thrips and application of the dsRNA. Specifically, the invention provides a dsRNA construct, the construct of the dsRNA is a double strand, and the construct of the dsRNA comprises a nucleotide sequence of an insect nymph and / or adult stage regulation related gene or fragment; wherein the insect nymph and / or adult stage regulation related gene is an Actin gene, and the dsRNA is obtained by amplifying a sequence as shown in SEQ ID NO: 9-10. The fusion type dsRNA is designed to target a plurality of conservative thysanoptera actin gene dsRNA, so that the multi-killing effect is achieved, and the prevention and treatment effect of the fusion type dsRNA is equivalent to that of spinetoram which is commonly used in the market at present.

The disclosure provides, for example, double stranded DNA (dsDNA) molecules comprising one or more chemically modified nucleobases. In some embodiments, the dsDNA molecule is circular and comprises a first strand and a second strand, wherein the first strand comprises one or more chemically modified nucleobases, and the second strand is free of chemically modified nucleobases. In some embodiments, the dsDNA molecule comprises a promoter sequence and an effector sequence that encodes an effector.

Methods of assessing double-stranded plasmid DNA isoforms to determine a proportion of supercoiled plasmid DNA compared to linear plasmid DNA, and / or open circle plasmid DNA are provided according to aspects of the present disclosure, which include: preferentially labelling double-stranded plasmid DNA; flowing the labeled double-stranded plasmid DNA through a polymeric separation medium in a microchannel into a detection region; detecting the detectable label to determine: a) an amount of time taken by the labeled plasmid DNA isoforms to flow through the polymeric separation medium in the microchannel into the detection region, and / or b) strength of the signal of the detectable label in the detection region; and comparing a) and / or b) to a reference standard thereby determining a proportion of supercoiled plasmid DNA compared to linear plasmid DNA, and / or open circle plasmid DNA in the fluid sample comprising plasmid DNA.

The invention relates to the field of veterinary drug residue detection, in particular to a fluorescence sensor based on DNA molecular machine mediated split Cas12a and application of the fluorescence sensor to ofloxacin detection, the fluorescence sensor comprises a system A, a system B and a system C. According to the final concentration of each system, the system A comprises an AP / CP double-stranded compound and a 20 mM Tris-HCl buffer solution 1, the system B comprises a double-stranded compound, an sRNA probe and a 10 mM Tris-HCl buffer solution 2, and the system C comprises a DNA molecular machine mediated split Cas12a. The system C comprises a Cas12a / hRNA compound, a pDNA / psG43 signal probe, ThT and a 20 mM Tris-HCl buffer solution; aP in the AP / CP double-chain compound is an aptamer of ofloxacin, and AP and CP in the AP / CP double-chain compound are paired and connected through complementary base pairing; the sequences of AP, CP, PS, SS, sRNA, hRNA, pDNA and psG43 are respectively as shown in SEQ ID NO.1-SEQ ID NO.8. The invention also discloses a kit for detecting the content of the protein. According to the application, the nucleic acid aptamer of ofloxacin is utilized, based on a DNA molecular machine and SCas12a as a signal amplification technology and sulfo-modified G43 as a fluorescent signal probe, even if the content of ofloxacin in a sample is extremely low, detection can be accurately realized.

The invention discloses an engineered artificial vesicle and application of the engineered artificial vesicle in multiple in-situ detection of urine exosome miRNA, and belongs to the field of biosensors. The method comprises the following steps: constructing planar framework nucleic acid simultaneously modified with cholesterol and an aptamer through annealing reaction; a molecular beacon with a fluorophore is designed according to a target gene sequence, and the molecular beacon and a double-strand specific nuclease (DSN) system are jointly encapsulated in an artificial vesicle; the engineered artificial vesicles and a sample to be detected are incubated, the fluorescence modified molecular beacons can specifically recognize target genes, cyclic cutting and signal amplification are achieved under DSN mediation, and the broken molecular beacons release fluorescence signals; all fluorescence signals in the vesicles are collected through a fluorescence imaging technology, and a characteristic fluorescence spectrum of the to-be-detected sample is obtained. According to the present invention, the miRNA heterogeneity analysis at the single vesicle level and the precise multi-target detection of bladder cancer and other diseases can be achieved, and the problems of low throughput, target number limitation and the like of the existing vesicle in-situ detection are effectively overcome.

The invention discloses a liver cancer diagnostic kit for sequencing by combining a DNA logic gate with an aptamer, belongs to the field of diagnostic kits, and particularly relates to an AND gate logic operation double strand consisting of DNA sequences of cT1, cT2, cT35 and I, and an amplification reagent containing S-II, W, F, F-Cy5, M1 and M1-Cy3 based on entropy driven amplification reaction. After the PBMC is extracted from whole blood, the kit can be used for logically analyzing whether target proteins corresponding to the aptamers Apt51, GR-30 and IBA exist on the PBMC at the same time or not, so that early screening of the liver cancer is realized. A specific target of a liver cancer patient is found through aptamer single-cell high-throughput sequencing, and the kit can effectively distinguish healthy people from cancer patients.

The invention relates to the technical field of biological detection, and discloses a method for double-ring detection of lung cancer L858R mutation based on CRISPR / Cas9 and DNA, and the method comprises the following steps: enriching circulating tumor cells CTC from a blood sample; extracting genomic DNA (deoxyribonucleic acid) of the enriched CTC; carrying out amplification on the DNA fragment containing the L858R mutation by adopting recombinase polymerase amplification RPA (recombinase polymerase amplification); the amplified DNA fragment is hybridized with a recognition ring in a DNA double-ring structure, a DNA double ring is composed of the recognition ring and a report ring, and a part of region of the recognition ring is hybridized with a corresponding fragment of the report ring to form a staggered double-chain region; when L858R mutation exists, the mutated DNA and a recognition ring are hybridized to activate a CRISPR / Cas9 system, and under the guidance of sgRNA, Cas9 cuts hybrid double strands and releases a report ring; carrying out rolling circle amplification RCA under the action of DNA polymerase by taking the released report ring as a template; a fluorescent probe is used for detecting an RCA amplification product, and whether L858R mutation exists or not is judged according to a fluorescence signal. The CRISPR / Cas9 system is combined with a DNA double-ring structure, so that the ultra-sensitive and specific detection on the lung cancer L858R mutation is realized.

The application provides a preparation of a sema7A monoclonal antibody 8A12 and a treatment effect of the sema7A monoclonal antibody 8A12 on lupus nephritis, wherein the CDR-H1 of the heavy chain variable region of the monoclonal antibody 8A12 is an amino acid sequence shown in SEQ ID No. 1, the CDR-H2 of the heavy chain variable region is an amino acid sequence shown in SEQ ID No. 2, and the CDR-H3 of the heavy chain variable region is an amino acid sequence shown in SEQ ID No. 3; the CDR-L1 of the light chain variable region of the monoclonal antibody 8A12 is an amino acid sequence shown in SEQ ID No. 4, the CDR-L2 of the light chain variable region is an amino acid sequence shown in SEQ ID No. 5, and the CDR-L3 of the light chain variable region is an amino acid sequence shown in SEQ ID No. 6. The monoclonal antibody 8A12 can effectively inhibit the expression up-regulation of IL-1beta, TNF-alpha and IL-6 caused by Sema7A recombinant protein, can effectively inhibit the macrophage inflammatory response induced by Sema7A, and can continuously and effectively reduce serum anti-double-stranded DNA antibodies and kidney damage of lupus mice. The monoclonal antibody 8A12 can be used for preparing a pharmaceutical composition for preventing and / or treating systemic lupus erythematosus and / or lupus nephritis thereof.

The present disclosure provides double stranded ribonucleic acid (dsRNA) agents for inhibiting expression of a target gene, comprising an antisense strand which is complementary to the target gene; a sense strand which is complementary to the antisense strand and forms a double stranded region with the antisense strand; at least one alpha-v-beta-6 (αvβ6) integrin targeting ligand that mediates delivery to muscle tissue conjugated to at least one strand; and at least one in vivo delivery enhancing moiety conjugated to one or more internal positions on at least one strand. The present disclosure also provides compositions comprising such dsRNA agents, and methods of use thereof for treating a subject having a disorder that would benefit from reduction in expression of the target gene.