476 results about "Double stranded" patented technology

The invention discloses a target segment for preventing and treating tomato brown fruit wrinkling virus in tobacco, double-stranded RNA (Ribonucleic Acid) and application of the target segment and the double-stranded RNA. Wherein the nucleotide sequence of the target segment is as shown in SEQ ID NO.3, or a sequence which is as shown in SEQ ID NO.3, has at least 90% of homology and has coded amino acid as shown in SEQ ID NO.4 is substituted. The invention also discloses a double-stranded RNA of a targeted interference target section, and the double-stranded RNA can significantly inhibit the proliferation of the tomato brown crinkled fruit virus in tobacco by injecting or smearing tobacco leaves, so that the double-stranded RNA can be effectively applied to prevention and control of tobacco diseases caused by the tomato brown crinkled fruit virus. The method for preventing and controlling the tomato brown crinkled fruit virus in the tobacco by using the double-stranded RNA is safe, harmless and efficient, and provides a new path for preventing and controlling plant virus diseases by using RNAi (Ribonucleic Acid Interference).

Disclosed are, inter alia, double stranded RNAi (dsRNAi) agents inhibiting expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), for example, human HMGCR, compositions including the same, and methods of treatment using the same.

A dsRNA and double-stranded RNAi agent for inhibiting the expression of INHBE and a use thereof. The dsRNA and the double-stranded RNAi agent can be used for inhibiting the expression of INHBE to prevent and treat a metabolic disorder (such as a metabolic syndrome).

The invention provides siRNA for targeted regulation and control of HBV gene expression and application of the siRNA. The double-stranded RNAi agent comprises an antisense strand and a sense strand complementary to the antisense strand forming a double-stranded region, and the nucleotide sequence of the antisense strand is as shown in SEQ ID NO: 8, or the nucleotide sequence of the antisense strand is a modified sequence of the sequence as shown in SEQ ID NO: 8. Cell and animal experiment results show that the double-stranded RNAi agent provided by the invention can significantly reduce the expression of one or more HBV genes and block the life cycle of viruses, and can be used for developing drugs for treating HBV gene expression related diseases.

The invention discloses a double-stranded RNA (Ribonucleic Acid) aiming at hemiptera insects as well as a pesticide composition and application of the double-stranded RNA. The dsRNA provided by the invention is used as a biopesticide, has the characteristics of high efficiency, specificity, fast degradation and no residue, realizes the prevention and control of brown planthopper through an RNAi mechanism, and is beneficial to reducing the use of chemical pesticides. The invention provides a brand new brown planthopper target gene as a target of RNA interference. The survival rate result shows that the genes play an important role in the survival process of the brown planthopper, are effective targets for preventing and treating the brown planthopper by using a gene silencing means, and have a wide application prospect. Correspondingly, the invention provides a plurality of dsRNA sequences aiming at the targets, and part of the sequences show unprecedented brown planthopper control efficiency.

The present invention relates to RNAi agents, e.g., double stranded RNA (dsRNA) agents, targeting the complement component C3 gene (C3). The invention also relates to methods of using such RNAi agents to inhibit expression of a C3 gene and to methods of preventing and treating a C3-associated disorder, e.g., cold agglutinin disease (CAD), warm autoimmune hemolytic anemia, and paroxysmal nocturnal hemoglobinuria (PNH), lupis nephritis (LN), bullous pemphigoid, Pemphigus, e.g., Pemphigus vulgaris (PV) and Pemphigus foliaceus (PF), and C3 glomerulopathy.

The technology described herein relates to 5'-modified nucleosides, nucleotides, oligonucleotides and double-stranded RNAs, e.g., siRNAs, and kits comprising them and methods of their use for inhibiting target genes.

The invention provides a method for delivering double-stranded RNA (Ribonucleic Acid) molecules into locusts through epidermis, a product and application. The method provided by the invention comprises the following steps: mixing double-stranded RNA molecules to be delivered with a polymer carrier as shown in a formula 1 to obtain a mixture of the double-stranded RNA molecules and the polymer carrier; adding a surfactant into the mixture of the double-stranded RNA molecules and the polymer carrier to obtain a delivery mixture; the delivery mixture is contacted with the epidermis of the locust, so that the double-stranded RNA molecule enters the locust body through the epidermis. According to the method provided by the invention, the double-stranded RNA molecules can penetrate through relatively thick epidermis to enter the locust body, so that the stability of the double-stranded RNA molecules in the locust body is improved, and efficient gene interference is realized. The method provided by the invention can be used for research of locust gene functions and application of RNA pesticides.

One aspect of the present invention relates to double-stranded RNA (dsRNA) agent capable of inhibiting the expression of a target gene. Other aspects of the invention relate to pharmaceutical compositions comprising these dsRNA molecules suitable for therapeutic use, and methods of inhibiting the expression of a target gene by administering these dsRNA molecules, e.g., for the treatment of various disease conditions.

The invention relates to the technical field of agricultural biology, and particularly provides a dsRNA nanocrystallization preparation for preventing and treating areca yellows as well as a preparation method and application of the dsRNA nanocrystallization preparation. Areca yellows are a destructive disease caused by areca symptomless virus type 1 (APV1), eight key target gene segments of an APV1 virus genome are screened, specific double-stranded RNA is designed, a nano-carrier is used for wrapping and delivering a dsRNA mixture obtained by transcription of the eight target gene segments, and the areca yellows are obtained by screening. And a nanocrystallization preparation for improving the environmental stability and RNA interference efficiency of the dsRNA is prepared. Experimental results show that the nanocrystallization preparation can effectively inhibit the key gene expression of APV1 virus, reduce the virus load and improve the resistance of areca catechu to yellows. The invention provides an accurate, efficient and environment-friendly biological prevention and control strategy, and has a wide agricultural application prospect.

The invention discloses dsRNA of a targeted potato beetle heat shock protein HSP70-KDa3 gene as well as a preparation method and application of the dsRNA of the targeted potato beetle heat shock protein HSP70-KDa3 gene. In order to solve the problems that an existing RNAi target gene is prone to off-target, poor in field stability and the like, the potato beetle heat shock protein gene LdHSP70-KDa3 serves as an RNA interference target for the first time, and a double-stranded RNA molecule shown as SEQ ID NO: 1 and a preparation method for preparing the double-stranded RNA are provided. Feeding experiments prove that the double-stranded RNA can efficiently silence target genes, the survival rate of potato beetle larvae is remarkably reduced, and feeding and growth development of the potato beetle larvae are inhibited. In addition, the invention further provides a biological preparation containing the double-stranded RNA and application of the biological preparation in prevention and treatment of potato beetles. The dsRNA targeting the HSP70-KDa3 gene provided by the invention has the characteristics of high efficiency, specificity, environmental friendliness and the like, and a new technical scheme is provided for green prevention and control of potato beetles.

The invention discloses a double-stranded RNA (Ribonucleic Acid) aiming at hemiptera insects as well as a pesticide composition and application of the double-stranded RNA. The dsRNA provided by the invention is used as a biopesticide, has the characteristics of high efficiency, specificity, fast degradation and no residue, realizes the prevention and control of brown planthopper through an RNAi mechanism, and is beneficial to reducing the use of chemical pesticides. The invention provides a brand new brown planthopper target gene as a target of RNA interference. The survival rate result shows that the genes play an important role in the survival process of the brown planthopper, are effective targets for preventing and treating the brown planthopper by using a gene silencing means, and have a wide application prospect. Correspondingly, the invention provides a plurality of dsRNA sequences aiming at the targets, and part of the sequences show unprecedented brown planthopper control efficiency.

The invention discloses a double-stranded RNA (Ribonucleic Acid) aiming at hemiptera insects as well as a pesticide composition and application of the double-stranded RNA. The dsRNA provided by the invention is used as a biopesticide, has the characteristics of high efficiency, specificity, fast degradation and no residue, realizes the prevention and control of brown planthopper through an RNAi mechanism, and is beneficial to reducing the use of chemical pesticides. The invention provides a brand new brown planthopper target gene as a target of RNA interference. The survival rate result shows that the genes play an important role in the survival process of the brown planthopper, are effective targets for preventing and treating the brown planthopper by using a gene silencing means, and have a wide application prospect. Correspondingly, the invention provides a plurality of dsRNA sequences aiming at the targets, and part of the sequences show unprecedented brown planthopper control efficiency.

Disclosed are, inter alia, therapeutic oligonucleotides for use in silencing a target gene in a cell or in a subject.

The invention discloses a target segment and double-stranded RNA for preventing and controlling propagation of tomato brown crinkled fruit virus through tomato seeds and application of the target segment and the double-stranded RNA. Wherein the nucleotide sequence of the target segment is as shown in SEQ ID NO.3, or a sequence which is as shown in SEQ ID NO.3, has at least 90% of homology and has coded amino acid as shown in SEQ ID NO.4 is substituted. The invention further discloses double-stranded RNA of the target interference target section, the double-stranded RNA is used for single isolation treatment or batch treatment of germinated tomato seeds, and the infection rate of the tomato brown crinkled fruit virus on germinated tomato seedlings can be effectively reduced under the condition that the germination rate of the tomato seeds is not affected. The method for preventing and controlling propagation of the tomato brown crinkled fruit virus through the tomato seeds by using the double-stranded RNA is safe and harmless, and has a good application prospect.

The invention discloses a method for site-specific replacement or insertion of DNA fragments in a plant genome, and belongs to the technical field of biology. The method disclosed by the invention comprises the following steps: editing a genome of a plant protoplast by adopting a CRISPR / Cas9 system and taking a double-stranded DNA repair template as a donor template to obtain an edited protoplast; and carrying out culture regeneration on the edited protoplast by utilizing a plant protoplast transformation method to obtain a site-directed mutagenesis plant in the genome so as to realize site-directed mutagenesis of the target plant genome. The method disclosed by the invention can be used for accurately replacing DNA in a plant genome at a fixed point. According to the invention, a recognition sequence of recombinase can be accurately inserted into a genome at a fixed point through homologous recombination, and then accurate fixed-point replacement of a gene large fragment is realized by virtue of a specific recombinase system.

A modified dsRNA, a double-stranded RNAi agent, and a use thereof. The modified dsRNA and the double-stranded RNAi agent can reduce non-specific binding, thereby reducing off-target toxicity, and improving the inhibitory activity on each target.

The invention provides a macro virus group sequencing library construction method for removing host ribosome RNA through an enzyme method and a kit, and belongs to the technical field of biology. The method comprises the following steps: hybridizing total RNA (Ribonucleic Acid) of a sample with a DNA (Deoxyribose Nucleic Acid) probe aiming at host ribosome RNA to form a DNA / RNA hybrid chain; rNase H is used for digesting RNA in the hybrid chain, and DNase I is used for digesting the remaining DNA probe in the system; purifying to obtain virus RNA (Ribonucleic Acid); carrying out fragmentation treatment on the virus RNA; carrying out reverse transcription on the fragmented RNA to synthesize a first chain cDNA, then synthesizing a second chain cDNA, and carrying out terminal repair and dA tail addition to obtain double-chain cDNA; connecting the linker with the dT tail with the double-chain cDNA; and carrying out PCR amplification and purification on the connection product to obtain the macro virus group sequencing library. According to the construction method and the kit, the problem that the full-length sequence of the virus cannot be obtained from a low-load sample can be solved.

The present invention provides a novel double-stranded siRNA, a conjugate thereof and a use thereof, wherein the double-stranded siRNA and the conjugate thereof can inhibit LAP gene expression, and can be used for preparing drugs for treating and / or preventing LAP-related diseases.

The invention provides dsRNA for preventing and controlling liriomyza sativae and a pesticide compound thereof, the pesticide compound is formed by mixing dsRNA and an SPc star-type cation nano-carrier according to the mass ratio of 1: 1, and a nano-platform is formed through self-assembly and is used for targeted delivery of double-stranded RNA (dsRNA). The compound provided by the invention can significantly reduce the degradation rate of dsRNA in a plant leaf surface environment, can penetrate through a plant leaf cuticle and enter mesophyll tissue, so that larvae latent in the leaf can intake the compound, and the dsRNA targeted to LtCht2 is designed to specifically cut LtCht2 mRNA through the RNA interference pathway effect in the larva body, so that chitin metabolism disorder is caused, and the expression of the LtCht2 in the larva body is inhibited. Therefore, normal pupation and eclosion of the larvae are blocked, and finally the purpose of controlling the population of the liriomyza clover is achieved.

The invention discloses a composition for preventing and treating eggplant root rot, and relates to the technical field of biological prevention and treatment. The composition consists of three parts, namely a double-stranded RNA (Ribonucleic Acid) delivery body controlled by a kaolinite nanotube-metal polyphenol network-boric acid ester, a nitric oxide / hydrogen sulfide double-pulse microcapsule and volatile organic compound particles of beta-cyclodextrin cross-linked aerogel, wherein the three parts are matched according to the total mass ratio of 1: (0.8-1.5): (0.8-1.2). Through structural design, time sequence release of non-overlapping peak values is realized, and a prevention and control mode for inhibiting environment enhancement host precise silencing is formed. The deliverer can be selectively disintegrated under the condition of pathogen-induced organic acid, iron chelating agent and hydrogen peroxide, so that the targeted release of the double-stranded RNA is realized; the double-pulse microcapsule respectively releases nitric oxide and hydrogen sulfide in a transplanting stress stage and an early disease stage, and induces host defensive gene expression; the aerogel particles continuously release 2, 3-butanediol and aromatic aldehyde to inhibit germination of pathogenic bacteria and serve as rhizosphere signals to enhance host resistance.

The present disclosure relates to RNAi agents, e.g., double stranded RNAi agents such as small interfering RNA (siRNA) molecules, able to inhibit proprotein convertase subtilisin kexin 9 (PCSK9) gene expression. Also disclosed are pharmaceutical compositions that include PCSK9 RNAi agents and methods of use thereof. The PCSK9 RNAi agents disclosed herein may be conjugated to targeting ligands, including ligands that comprise N-acetyl-galactosamine, to facilitate the delivery to hepatocyte cells. Delivery of the PCSK9 RNAi agents in vivo provides for in vivo provides for inhibition of PCSK9 gene expression and thereby reduction of PCSK9 protein. The RNAi agents can be used in methods of treatment of diseases or disorders mediated at least in part by PCSK9 gene expression, including among others hypercholesterolemia, familial hypercholesterolemia including heterozygous familial hypercholesterolemia (HeFH) and homozygous familial hypercholesterolemia (HoFH), familial hypobetalipoproteinemia, hyperlipidemia, coronary artery disease, polygenic dyslipidemia, heart disease, cardiovascular disease (CVD) including clinical atherosclerotic cardiovascular disease (ASCVD).

Provided are T7 RNA polymerase variants and the use thereof. The T7 RNA polymerase variants may reduce dsRNA byproducts produced, e. g. during an IVT reaction, while retaining RNAP activity.

The invention provides a preparation method of a pH-responsive release nucleic acid delivery system, and belongs to the technical field of nucleic acid delivery systems.The preparation method comprises the steps that 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride is used for enhancing the amino density of the surface of bovine serum albumin, modified bovine serum albumin and double-stranded ribonucleic acid are mixed according to the mass ratio of 10: 1, and a mixture is obtained; the preparation method comprises the following steps: adding glutaraldehyde as a cross-linking agent under the condition that the ratio of water to ethanol is 3: 1 to form Schiff base, reacting at 4 DEG C for 4-5 hours, and then performing ultrafiltration purification to obtain the double-stranded ribonucleic acid modified bovine serum albumin nanoparticles, thereby solving the technical problem of poor stability of double-stranded ribonucleic acid in noctuidae insects. And an efficient and accurate technical solution is provided for agricultural pest control.

The invention relates to a dsRNA insecticide for preventing and treating thrips and a preparation method of the dsRNA insecticide. Belongs to the field of biotechnology and agricultural application. The dsRNA insecticide for preventing and treating the thrips is a preparation taking dsRNA as an effective component, and the dsRNA is of a double-stranded RNA structure containing a positive-sense strand and an antisense strand; the nucleotide sequence of the positive-sense strand is as shown in SEQ ID NO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9 or SEQ ID NO. 11. The invention proves that dsACT and dsVATB can cause degradation of messenger RNA (Ribonucleic Acid) of ATP (Adenosine Triphosphate) enzyme coding genes in thrips and reduce the expression level of related genes; laboratory test and field efficacy test results jointly show that after the dsRNA insecticide is sprayed, the growth and development of the thrips are affected, individual death is further caused, and the achievement lays a good theoretical and application foundation for prevention and treatment of the thrips.

The invention discloses a compound composition of fluopyram and dsRNA (double-stranded ribonucleic acid) and application of the compound composition in prevention and control of nematodes. The method comprises the following steps: soaking pine wood nematodes by adopting a fluopyram medicament with LC20 concentration, collecting samples at different time points respectively, and carrying out transcriptome analysis. The pesticide compound composition for effectively poisoning and killing the pine wood nematodes is obtained by comparing gene expression differences at different time points, screening out remarkably up-regulated detoxification genes BxCYP, BxCES and BxUDP, designing and synthesizing double-stranded RNA (ds BxCYP, ds BxCES and ds BxUDP) according to the detoxification genes, and compounding the double-stranded RNA with fluopyram. The pesticide compound composition has a remarkable synergistic prevention and control effect on pine wood nematodes, is expected to become a novel and efficient pine wood nematode prevention and control agent, and provides a new strategy and theoretical basis for prevention and control of pine wood nematode diseases.

One aspect of the present invention relates to double-stranded RNA (dsRNA) agent capable of inhibiting the expression of a target gene. Other aspects of the invention relate to pharmaceutical compositions comprising these dsRNA molecules suitable for therapeutic use, and methods of inhibiting the expression of a target gene by administering these dsRNA molecules, e.g., for the treatment of various disease conditions.

Provided are compositions, systems and methods for removal of double stranded RNA from a mixture utilizing affinity agents having high binding affinity and selectivity for double stranded RNA over single stranded RNA.

Disclosed herein are double stranded ribonucleic acids (dsRNAs) and synthetic antisense oligonucleotides (ASOs) for use in inhibiting the expression of MUC5B and in the treatment of a subject having a disease or disorder characterized by overexpression of MUC5B, including in subjects having lung disease or in a subject at risk of developing lung disease, for example, in a subject having or at risk of developing idiopathic pulmonary fibrosis.

This invention discloses a method for directed evolution of antibiotic-resistant Escherichia coli strains based on cytidine deaminase, belonging to the field of microbial directed evolution and genetic engineering technology. This method uses E. coli as the host, introducing a recombinant plasmid expressing an optimized double-stranded cytidine deaminase mutant. Utilizing the low toxicity and high mutagenicity of this mutant, continuous passage evolution is carried out under gradient concentrations of aminoglycoside antibiotics (kanamycin and streptomycin). Combined with whole-genome sequencing, molecular docking, and reverse genetics verification, the A145T missense mutation in the wcaE gene is identified as the core functional site. This invention overcomes the shortcomings of traditional spontaneous and chemical mutagenesis, which suffer from low efficiency and significant strain damage. The mutation type is controllable, the evolutionary cycle is short, and the obtained engineered strains can tolerate up to 300 mg / L kanamycin while exhibiting streptomycin cross-resistance, and the genetic stability of the tolerance trait is strong. This method is simple, highly reproducible, and suitable for industrial breeding of stress-resistant E. coli, and can be widely applied in antibiotic fermentation, industrial microbial culture, and other scenarios.