392 results about "Double stranded" patented technology

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.

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.

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 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 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).

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.

The present disclosure provides methods and tools that allow for the production of large quantities of double-stranded RNA (dsRNA) in bacterial cells, including plasmid vectors that can be used to transform Gram-positive or Gram-negative bacterial cells, such that the cells produce high yields of target dsRNA.

Provided are a double-stranded ribonucleic acid (dsRNA) for inhibiting C5 gene expression, a double-stranded RNAi agent, and a use thereof. The dsRNA and the double-stranded RNAi agent can be used for inhibiting C5 gene expression, thereby preventing and treating diseases and / or conditions related to C5 gene expression.

The present application relates to siRNA and oligonucleotide agents for use in the prevention or treatment of SOD1-related neurodegenerative diseases or conditions, such as amyotrophic lateral sclerosis, ALS. The oligonucleotide agent comprises a double-stranded targeting oligonucleotide (siRNA) and a non-targeting single-stranded oligonucleotide (ACO), wherein the siRNA targets the mRNA region of the target gene SOD1.

The application discloses two kinds of small RNA compositions for improving biological control effect of termites, and microRNA-like RNA (milRNA) sequences are milR-32 and milR-48 respectively. Double-stranded RNA can enhance the effect of the beetle green fungus on killing termites. The milR-32 and the milR-48 belong to the milRNA coded by the green fungus, and the gene sequences are derived from the transcriptome database of the beetle green fungus. The milR-32 and the milR-48 mimics are designed and synthesized based on the milR-32 and the milR-48 sequences respectively. After the beetle green fungus is used to infect the termites taking the milR-32 or the milR-48 mimic, the infection mortality of the termites is significantly increased. The mimic designed based on the milR-32 and the milR-48 sequences can significantly improve the biological control effect of the termites.

To provide dsRNA molecules which are advantageous for inhibition of target gene expression while having reduced off-target gene silencing effects.SOLUTION: The present invention relates to double-stranded RNA (dsRNA) agent capable of inhibiting the expression of a target gene. The antisense strand of the dsRNA molecule comprises at least one thermally destabilizing nucleotide occurring at a seed region; the dsRNA comprises at least four 2'-fluoro modifications; and the sense strand of the dsRNA molecule comprises a ligand, where the ligand is an ASGPR ligand.SELECTED DRAWING: None

The present invention provides technology for inhibiting or suppressing proliferation of cells. Disclosed herein is a double-stranded RNA that has a first strand and a second strand which is complementary to the first strand, wherein the first strand has a main sequence which comprises 19-23 bases and in which a base at the 5'-end is guanine (G) or cytosine (C) and an additional sequence which comprises 2-4 bases and which is added to the 3'-end side of the main sequence. The main sequence is a portion of a base sequence that encodes semaphorin-3A, the portion including at least a portion of a base sequence that encodes a signal peptide region of the semaphorin-3A.

The invention belongs to the technical field of bioengineering, and particularly relates to a method for purifying single-stranded DNA and application thereof. The invention provides a method for purifying single-stranded DNA (deoxyribonucleic acid). The method comprises the following steps: S1, carrying out polymerization reaction on a primer of which the 5'end is modified with an acrylamide group to generate a linear polyacrylamide modified primer LPA-primer; s2, by taking the LPA-primer as a primer, carrying out PCR (Polymerase Chain Reaction) amplification, so as to obtain a double-stranded DNA (Deoxyribose Nucleic Acid) NALPA-dsDNA (Deoxyribose Nucleic Acid) crosslinked by polyacrylamide; s3, carrying out denatured agarose gel electrophoresis on the LPA-dsDNA under an alkaline condition, and separating a target ssDNA from a non-target ssDNA by utilizing a mobility difference caused by a polyacrylamide modified group; and S4, carrying out gel cutting and recycling on the target ssDNA strip to obtain the purified ssDNA. The result of the embodiment shows that the method can effectively improve the purity of the single-stranded DNA and reduce the cost.

This invention discloses five RNAi molecules targeting the genome of the small cutworm, all of which are double-stranded RNA molecules composed of a sense strand and a complementary antisense strand, wherein the nucleotide sequence of the sense strand is selected from SEQ ID NOs:1-5. The lethality of these RNAi molecules against the small cutworm is all above 80%.

The invention belongs to the technical field of biological medicine, and particularly relates to a rabies virus G protein carrier, a rabies virus vaccine and a preparation method and application of the rabies virus G protein carrier and the rabies virus vaccine. The rabies virus G protein vector pscAAV-G is obtained by inserting a coding gene of rabies virus G protein into a coding region between inverted terminal repetitive sequences at two ends of a double-chain adeno-associated virus vector, so that the problem of expression delay of single-chain AAV (ssAAV) is solved, and rapid and efficient expression of the rabies virus G protein is realized; furthermore, an endogenous nucleic acid adjuvant is embedded into a non-coding region of the pscAAV-G, so that the rabies virus G protein carrier pscAAV-G-Adj is prepared, and Th1 type immune response is remarkably improved. According to a rabies virus vaccine prepared by using the rabies virus G protein carrier, the antigen expression efficiency and immunogenicity of an AAV carrier vaccine are remarkably improved, long-acting protection of single inoculation is realized, and the defects that an existing rabies vaccine is short in immune cycle and needs to be inoculated for multiple times are overcome.

The invention discloses a method for regulating and controlling migratory locust embryo diapause by a BTB transcription factor LOCMI00260 and application of the method. According to the present invention, the full length of the BTB transcription factor LOCMI00260 is cloned from migratory locust, the specific double-stranded RNA interference sequence fragment dsRNA of the BTB transcription factor LOCMI00260 is developed, and the dsRNA of the BTB transcription factor LOCMI00260 is introduced into the migratory locust egg by using the microinjection method; the result shows that the migratory locust BTB transcription factor LOCMI00260 can improve the diapause rate of migratory locust. The invention provides a new means and thought for migratory locust prevention and treatment.

The present disclosure relates to RNAi agents, e.g., double-stranded RNAi agents, e.g., siRNAs, capable of inhibiting expression of the subunit beta E (INHBE) gene. Also disclosed are pharmaceutical compositions comprising the INHBE RNAi agents, and methods of use thereof. The INHBE RNAi agents disclosed herein can be conjugated to targeting ligands to facilitate delivery to cells, including to hepatocytes. Delivery of the INHBE RNAi agent in vivo results in inhibition of expression of the INHBE gene. RNAi agents may be used in methods of treating diseases, disorders, or conditions mediated in part by the expression of the INHBE gene, such as obesity, diabetes, liver inflammation, dyslipidemia, or metabolic diseases.