266 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 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 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).

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.

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 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.

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 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.

Provided are double-stranded RNAs for inhibiting the expression of angiotensinogen (AGT) in a cell, vectors and cells comprising their encoding nucleotides, and methods of using the dsRNAs, vectors or cells to treat diseases or symptoms mediated or associated with the expression of AGT in a subject.

This invention discloses a nanoRNA delivery system and its application in the control of Sclerotinia sclerotinia disease, relating to the field of biotechnology. The preparation method of the nanoRNA delivery system includes the following steps: mixing chitosan, sucrose, and aspartic acid uniformly and then subjecting them to a hydrothermal reaction to obtain a quantum dot-chitosan degradation product complex; mixing the quantum dot-chitosan degradation product complex with double-stranded RNA to obtain the nanoRNA delivery system; the double-stranded RNA is used to silence Sclerotinia sclerotinia virulence-related transcription factor genes. The nanoRNA delivery system provided by this invention can effectively protect RNA and exhibits a very high inhibitory effect on Sclerotinia sclerotinia in plants, providing a novel technical solution for the green and precise control of Sclerotinia sclerotinia disease.

The invention relates to a method of treating ocular amyloidosis by reducing TTR expression in a subject by administering a double-stranded ribonucleic acid (dsRNA) that targets a TTR gene to the retinal pigment epithelium of the subject.

The present disclosure relates to RNAi agents, e.g., double stranded RNAi agents, able to inhibit xanthine dehydrogenase (XDH) gene expression. Also disclosed are pharmaceutical compositions that include XDH RNAi agents and methods of use thereof. The XDH RNAi agents disclosed herein may be conjugated to targeting ligands to facilitate the delivery to cells, including to hepatocytes. Delivery of the XDH RNAi agents in vivo provides for inhibition of XDH gene expression. The RNAi agents can be used in methods of treatment of diseases, disorders, or symptoms mediated in part by XDH gene expression, such as gout and hyperuricemia.

Disclosed is a method for purifying single-stranded RNA (ssRNA). More specifically disclosed is a method of purifying an ssRNA-containing sample containing double-stranded RNA (dsRNA) as an impurity using two different chromatography steps. The method includes steps of: (a) subjecting a sample containing ssRNA and containing double-stranded RNA (dsRNA) as an impurity to primary purification using affinity chromatography; and (b) subjecting the sample to secondary purification using at least one chromatography selected from the group consisting of size-exclusion chromatography (SEC), anion-exchange chromatography (AEX), and hydrophobic interaction chromatography (HIC). The method has a high mRNA recovery and a dsRNA removal rate of 95% or more, and is useful because it can minimize or appropriately control the dsRNA content in an mRNA sample, so that the mRNA can be developed into a drug regardless of the type of sequence, and can also maximize patient safety and drug efficacy.

The invention relates to double-stranded ribonucleic acid (dsRNA) compositions targeting the ANGPTL3 gene, as well as methods of inhibiting expression of ANGPTL3 and methods of treating subjects having a disorder of lipid metabolism, such as hyperlipidemia or hypertriglyceridemia, using such dsRNA compositions.

The invention provides an in-vitro transcription method based on RNA polymerase and a kit, which are suitable for the fields of mRNA synthesis, gene research and biological pharmacy. According to the method, the problems of low efficiency, poor stability and the like of traditional in-vitro transcription are solved through an optimization technology. The method comprises the following specific steps: providing a linear DNA template containing a T7 promoter; according to the method, site-directed mutagenesis or fusion protein modified high-activity T7RNA polymerase is combined with chemically modified NTPs and a spermine-BSA compound stabilizer to react under the conditions that the temperature is 37-42 DEG C and the pH is 7.5-8.5, and formation of double-stranded RNA is promoted through dynamic temperature control. A reaction system contains Mg < 2 + > / Mn < 2 + > and DTT, and enzyme activity and product integrity are optimized. The kit comprises an improved reagent and a standardized process, the transcription efficiency is improved by 2-5 times, the product yield is greater than or equal to 80%, and the activity is not lost after the kit is stored at 4 DEG C for 1 month. The method has the technical advantages that NTPs degradation resistance is achieved, template preparation is simplified, universality is high, the method is suitable for large-scale production of mRNA vaccines, gene therapy and the like, the production cost is remarkably reduced, and wide application of the in-vitro transcription technology is promoted.

Provided are a double-stranded RNA for inhibiting the expression of an INHBE gene, a cell containing same, and a method for treating diseases or symptoms mediated by or associated with INHBE in a subject using the dsRNA or cell.

This disclosure provides siRNAs that target and regulate HBV gene expression and their applications. The double-stranded RNAi agent comprises an antisense strand and a sense strand complementary to the antisense strand forming the double-stranded region. The nucleotide sequence of the antisense strand is shown in SEQ ID NO:13, or the nucleotide sequence of the antisense strand is a modified sequence of the sequence shown in SEQ ID NO:13. Cellular and animal experimental results show that the double-stranded RNAi agent provided in this disclosure can significantly reduce the expression of one or more HBV genes, block the viral life cycle, and can be used to develop drugs for treating HBV gene expression-related diseases.

This provides a dsRNA for inhibiting the expression of trans tyretin. [Solution] Disclosed are a double-stranded ribonucleic acid (dsRNA) that targets the trans tiretin (TTR) gene, and a method of using the dsRNA to inhibit TTR expression.

One aspect of the present invention relates to a double-stranded RNA (dsRNA) agent for modulating the expression of a target gene in the central nervous system (CNS), comprising: an antisense strand complementary to the target gene in the CNS; a sense strand complementary to the antisense strand; and one or more saturated or unsaturated C RNAs conjugated onto at least one strand via a linker or carrier as appropriate. 22 The present invention relates to a dsRNA agent comprising one or more lipophilic moieties containing a hydrocarbon chain. Another aspect of the present invention relates to a pharmaceutical composition comprising a dsRNA agent. Another aspect of the present invention relates to a method for modulating the expression of a target gene in CNS cell genes and a method for treating or preventing CNS damage in a subject, comprising administering a therapeutically effective amount of a dsRNA agent to cells or a subject.