147 results about "Single-Stranded RNA" patented technology

The present invention is based on the development of a dual promoter system (preferably a RNA pol I-pol II system) for the efficient intracellular synthesis of viral RNA. The resultant minimal plasmid-based system may be used to synthesize any RNA virus, preferably viruses with a negative single stranded RNA genome. The viral product of the system is produced when the plasmids of the system are introduced into a suitable host cell. One application of the system is production of attenuated, reassortant influenza viruses for use as antigens in vaccines. The reassortant viruses generated by cotransfection of plasmids may comprise genes encoding the surface glycoproteins hemagglutinin and neuramimidase from an influenza virus currently infecting the population and the internal genes from an attenuated influenza virus. An advantageous property of the present invention is its versatility; the system may be quickly and easily adapted to synthesize an attenuated version of any RNA virus. Attenuated or inactivated RNA viruses produced by the present invention may be administered to a patient in need of vaccination by any of several routes including intranasally or intramuscularly.

The present invention features compositions, methods, and kits useful in the treatment of viral diseases. In certain embodiments, the viral disease is caused by a single stranded RNA virus, a flaviviridae virus, or a hepatic virus. In particular embodiments, the viral disease is viral hepatitis (e.g., hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E). Also featured are screening methods for identification of novel compounds that may be used to treat a viral disease.

The present invention features compositions, methods, and kits useful in the treatment of viral diseases. In certain embodiments, the viral disease is caused by a single stranded RNA virus, a flaviviridae virus, or a hepatic virus. In particular embodiments, the viral disease is viral hepatitis (e.g., hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E) and the agent or combination of agents includes sertraline, a sertraline analog, UK-416244, or a UK-416244 analog. Also featured are screening methods for identification of novel compounds that may be used to treat a viral disease.

Double-stranded and single-stranded RNA molecules, and their use in methods for inducing interferon are provided. The interferon induction provides anti-viral and other medically useful effects, such as anti-cancer effects. Also provided are methods for reducing or inhibiting interferon induction exhibited by such molecules, particularly siRNA and shRNA molecules produced in vitro.

The invention describes composition and methods of use for novel hairpin blocked-cleavable primers. In one embodiment unblocking occurs through action of RNase H2. The method improves the specificity of PCR and reduces primer dimer events, enabling higher level multiplex reactions. Additionally, the invention protects RNA-containing primers from attack by single-strand RNases.

The present invention is based on the development of a dual promoter system (preferably a RNA pol I-pol II system) for the efficient intracellular synthesis of viral RNA. The resultant minimal plasmid-based system may be used to synthesize any RNA virus, preferably viruses with a negative single stranded RNA genome. The viral product of the system is produced when the plasmids of the system are introduced into a suitable host cell. One application of the system is production of attenuated, reassortant influenza viruses for use as antigens in vaccines. The reassortant viruses generated by cotransfection of plasmids may comprise genes encoding the surface glycoproteins hemagglutinin and neuraminidase from an influenza virus currently infecting the population and the internal genes from an attenuated influenza virus. An advantageous property of the present invention is its versatility; the system may be quickly and easily adapted to synthesize an attenuated version of any RNA virus. Attenuated or inactivated RNA viruses produced by the present invention may be administered to a patient in need of vaccination by any of several routes including intranasally or intramuscularly.

Double-stranded cDNA was synthesized from nucleic acid extracted from Norwalk virus purified from stool specimens of volunteers. One clone was isolated from a cDNA library constructed in a pUC-13 vector after amplification of the cDNA. The specificity of this cDNA (pUCNV-953) was shown by hybridization assays. The cDNA reacted with post (but not pre-) infection stool samples from Norwalk volunteers and with highly purified Norwalk virus, but not with other common enteric viruses such as hepatitis A virus and rotavirus. Finally, the probe detected virus in the same fractions of CsCl gradients in which viral antigen was detected using a specific Norwalk virus radioimmunoassay, and particles were detected by immune electron microscopy. Single-stranded RNA probes derived from the DNA clone after subcloning into an in vitro transcription vector were also used to show that the Norwalk virus contains a ssRNA genome of about 8 kb in size. The original clone was also used to detect additional cDNAs which represent at least 7 kb of nucleic acid of the Norwalk genome. The availability of a Norwalk-specific cDNA and the first partial genome sequence information allow rapid cloning of the entire genome and of establishment of sensitive diagnostic assays. Such assays can be based on detection of Norwalk virus nucleic acid or Norwalk viral antigen using polyclonal or monoclonal antibodies to proteins expressed from the cDNA or to synthetic peptides made based on the knowledge of the genome sequence. Assays using proteins deduced from the Norwlk virus genome and produced in expression systmes can measure antibody responses. Vaccines made by recombinant DNA technology are now feasible.

An antigen detection kit and an antigen detection method using the same are provided. The antigen detection kit comprises a capture antibody, a detection antibody bound to a single stranded DNA oligonucleotide, a single stranded RNA oligonucleotide complementary sequence to the DNA oligonucleotide, and an RNase.

Hepatitis C virus (HCV) is a major cause of chronic liver disease and affects over 270 million individuals worldwide. The HCV genome is a single-stranded RNA that functions as both a messenger RNA and replication template, making it an attractive target for the study of RNA interference. Double-stranded short interfering RNA (siRNA) molecules designed to target the HCV genome are disclosed herein.

The invention relates to the field of virology. The invention provides an isolated essentially mammalian negative-sense single stranded RNA virus (MPV) within the sub-family Pneumovirinae of the family Paramyxoviridae and identifiable as phylogenetically corresponding to the genus Metapneumovirus and components thereof.

A method of preparing normalized and / or subtracted cDNA; a method in which the cDNA that is normalized and / or subtracted is in the form of uncloned cDNA (cDNA tester); a method of preparing normalized and / or subtracted cDNA comprising the steps of: (a) preparing cDNA tester; (b) preparing normalization and / or subtraction RNA driver; (c) conducting normalization and / or subtraction in two steps in any order, or conducting normalization / subtraction as a single step and mixing the normalization / subtraction RNA driver with said cDNA tester; (d) adding an enzyme capable of cleaving single strand sites on RNA drivers nonspecifically bound to cDNA tester; (e) removing said single strand RNA driver cleaved in step d) from the tester and removing tester / driver hybrids; and (f) recovering the normalized and / or subtracted cDNA; and a method of efficiently preparing normalized and / or subtracted long-chain, full-coding, and full-length cDNA libraries are provided.

An antigen detection kit and an antigen detection method using the same are provided. The antigen detection kit comprises a capture antibody, a detection antibody bound to a single stranded DNA oligonucleotide, a single stranded RNA oligonucleotide complementary sequence to the DNA oligonucleotide, and an RNase.

The present invention provides new methods, employing a nucleotide integrase, for cleaving single-stranded RNA substrates, single-stranded DNA substrates, and double- stranded DNA substrates at specific sites and for inserting a nucleic acid molecule into the cleaved substrate. One method uses a nucleotide integrase to cleave one strand of a double-stranded DNA substrate. The method comprises the steps of: providing a nucleotide integrase comprising a group II intron RNA having two hybridizing sequences that are capable of hybridizing with two intron RNA binding sequences on the one strand of the DNA substrate, and a group II-intron encoded protein which binds to a first sequence element of the substrate; and reacting the nucleotide integrase with the double-stranded DNA substrate under conditions that permit the nucleotide integrase to cleave the one strand of the DNA substrate and to insert the group II intron RNA into the cleavage site. The method of cleaving both strands of a double-stranded DNA substrate comprises the steps of: providing a nucleotide integrase comprising a group II intron RNA having two hybridizing sequences that are capable of hybridizing with two intron RNA binding sequences on one strand of the substrate, and a group II-intron encoded protein that is capable of binding to a first sequence element and to a second sequence element in the recognition site of the substrate; and reacting the nucleotide integrase with the double-stranded DNA substrate such that the nucleotide integrase cleaves both strands of the DNA substrate and inserts the group II intron RNA into the cleavage site of the one strand. The method for cleaving a single-stranded nucleic acid substrate comprises the steps of: providing a nucleotide integrase having two hybridizing sequences that are capable of hybridizing with two intron RNA-binding sequences on the single-stranded substrate, and a group II intron encoded protein; and reacting the nucleotide integrase with the single stranded nucleic acid substrate for a time and at a temperature sufficient to allow the nucleotide integrase to cleave the substrate and to attach the group II intron RNA molecule thereto.

The present invention relates to compounds that modulate the replication of negative-sense, single-stranded RNA viruses, such as influenza virus, and the use of such compounds. The invention relates to methods for increasing the titer of negative-sense, single-stranded RNA viruses, such as influenza virus, in substrates for virus propagation (e.g., tissue culture). The invention also relates to the use of compounds that decrease virus replication as antiviral agents. The invention further relates to methods for identifying compounds that modulate the replication of negative-sense, single-stranded RNA viruses, in particular, influenza virus.

A new CRISPR-associated (Cas) protein, termed “CasM,” is described, as well as polynucleotides encoding the same and methods of using CasM for site-specific genome engineering. CasM proteins are capable of targeting and cleaving single-stranded RNA.

A method and a kit for detecting a target protein in a sample with a signal amplification strategy are provided. The signal amplification strategy is established for the aptamer-based molecular recognition of a target protein with concomitant release of single-stranded DNA (G-DNA), which binds complementarily to a single-stranded RNA comprising a fluorophore and a quencher (“F-RNA-Q”). The fluorescence-quenched RNA is then degraded by RNase H to result in a fluorescence signal, and the undamaged G-DNA is recycled to yield fluorescence amplification.

The invention relates to methods and compositions for modulating viral replication through double-stranded RNA-mediated gene silencing (RNAi), wherein the antiviral methods and compositions preferentially target opposite strand replication intermediates of single-stranded RNA viruses.

It is an object of the present invention to provide a novel molecule for antisense therapies which is not susceptible to nuclease degradation in vivo and has a high binding affinity and specificity for the target mRNAs and which can efficiently regulate expression of specific genes. The novel artificial nucleoside of the present invention has an amide bond introduced into a bridge structure of 2′,4′-BNA / LNA. The oligonucleotide containing the 2′,4′-bridged artificial nucleotide has a binding affinity for a single-stranded RNA comparable to known 2′,4′-BNA / LNA and has an increased nuclease resistance over LNA. Particularly, it is expected to be applied to nucleic acid drugs because of its much stronger binding affinity for single-stranded RNAs than S-oligo's affinity

The invention relates to small single stranded RNAs and analogs thereof (collectively “piRNA” herein), compositions comprising such piRNAs, and their uses in regulating target gene expression or as markers for certain disease states.

The invention relates to a fast detection method of nucleic acid of A H1N1 influenza virus, and a kit thereof, belonging to the technical field biotechnology. The invention provides a fast and synthermal RNA amplification method of A H1N1 influenza virus, and the accomplishment of the entire reaction depends on the cooperation of the AMV reverse transcriptase, the T7RNA polymerase and nuclease H (RNaseH), without the needs of the special instruments and the temperature cycling, the amplification efficiency is larger than or equal to that of the PCR, and the reaction product is single stranded RNA. The invention has the advantages of having equal or higher sensibility than that of the RT-PCR detection method, avoiding the pollution problem of the RT-PCR amplification product due to that the amplification product is RNA, finishing the reaction in one hour, and simple and convenient operation. The reaction can be monitored in real time and detected by the end-point method, can be used as the important tool in the detection of A H1N1 influenza virus.

Methods of making RNA duplexes and single-stranded RNAs of a desired length and sequence based on cleavage of RNA molecules at a defined position, most preferably with the use of deoxyribozymes. Novel deoxyribozymes capable of cleaving RNAs including a leader sequence at a site 3' to the leader sequence are also described.

A process for replicating or for replicating and expressing a sequence of interest in a plant, comprising: (i) an RNA replicon or a precursor thereof, said RNA replicon being derived from a plus-sense single stranded RNA virus and comprising at least one sequence of interest; and (ii) a helper replicon, or a precursor thereof, wherein said helper replicon is (a) incapable of systemic movement in said plant both in the presence and in the absence of said RNA replicon (i) and (b) capable of expressing in a plant one or more proteins necessary for systemic movement of said RNA replicon (i), whereby said RNA replicon (i) is capable of replicating or replicating and expressing said sequence of interest in said plant, but unable to move systemically in said plant in the absence of said one or more proteins expressed by said helper replicon (ii).

A system implemented on a computer is disclosed for automatically identifying strains of partial or complete capsid sequences of picoma and caliciviruses, two of the most highly diverse ssRNA virus families.

The invention discloses a RNA aptamer of a targeting hepatocyte and a nucleotide sequence thereof, and relates to the field of hepatic disease diagnosis and treatment. The RNA aptamer provides a novel specific and efficient molecule of a targeting hepatocyte for the field of hepatic disease diagnosis and treatment. In the invention, through using the new combinatorial chemical technology SELEX, and taking hepatic ASGPR (asialoglycoprotein receptor) large subunits as target proteins, an RNA aptamer which can specify the ASGPR is screened from a single-stranded RNA random library. The aptamer can be formed into a special stemloop structure in a random sequence region thereof, combines with big liver ASGP receptor subunit H1 protein with high specific affinity and can combine to liver cells in a targeted mode by high specific affinity. The RNA aptamer provides a new choice for developing a liver disease targeted diagnostic reagent and treatment drug.

Provided herein are methods and systems for structure-assisted evolution of multivalent aptamers using a single stranded DNA or single-stranded RNA nanostructure as a structural support. Also provided herein are methods for constructing ssDNA or ssRNA nanostructures as structural supports suitable for structure-assisted evolution of multivalent aptamers.

The invention relates to antisense oligonucleotides that are capable of bringing about specific editing of a target nucleotide (adenosine) in a target RNA sequence in a eukaryotic cell, wherein said oligonucleotide does not, in itself, form an intramolecular hairpin or stem-loop structure, and wherein said oligonucleotide comprises a non-complementary nucleotide in a position opposite to the nucleotide to be edited in the target RNA sequence.

The invention discloses a shear locus mediated by RNA induced silencing complex and application thereof. The locus can be applied in designing a factitial tiny RNA. In a DNA molecule coding a factitial tiny RNA precursor, the factitial tiny RNA precursor is a single-chain RNA capable of generating a tiny RNA in plant; the single-chain RNA forms a stem-loop structure; a nucleotide sequence containing sequence 1 in a sequence list is arranged in a fragment of the stem part of the stem-loop structure; two base pairs which are not complementary are arranged on the stem part of the stem-loop structure; and the two uncomplementary base pairs correspond to the ninth position and the tenth position from tail end of 5' in the sequence 1. The invention also discloses a method for culturing plant resisting cucumber mosaic virus. With the DNA molecule coding the factitial tiny RNA precursor, the plant with high efficient virus resistance can be obtained, so that the method is a biological engineering method with economy and efficiency.

The invention discloses a double-labelling fluorescence activation type probe and a method for detecting DNA by adopting the same. The probe is a single stranded RNA which is complementary with a basic group of to-be-detected DNA and has the length of 5-40 basic groups or is a single stranded DNA which is complementary with a basic group of to-be-detected DNA and has the length of 5-40 basic groups; 1 / 5-4 / 5 of basic groups in the DNA sequence are RNA basic groups; The two ends of the probe are labeled with a fluorophore and a quenching group. The probe is simple in structure and fluorescence is completely quenched; when the probe hybridizes with the to-be-detected DNA to form DNA / RNA heteroduplex, RNase H hydrolyzes an RNA chain in a DNA / RNA structure in a specificity manner to enable the fluorophore to be away from the quenching group and enable a fluorescence signal to be recovered; simultaneously, the dissociative probe further hybridizes with the to-be-detected DNA, is hydrolyzed by RNase H and generates the fluorescence signal; circulation is performed in such a way, so that the system fluorescence signal is remarkably amplified to realize rapid and high-sensitive detection of the to-be-detected DNA. The probe can be used for quantitative detection of DNA in an amplification product.