78 results about "Viral rna" patented technology

This invention generally relates to a novel RNA / mRNA production and amplification method using viral RNA replicase and / or RNA-dependent RNA polymerase (RdRp) enzymes as well as the associated mRNAs thereof. The present invention can be used for manufacturing and amplifying all varieties of RNA / mRNA sequences carrying at least an RdRp-binding site in the 5′- or 3′-end, or both. The RNA / mRNA so obtained is useful for not only producing mRNA vaccines and / or RNA-based medicines but also for generating the mRNA-associated proteins, peptides, and / or antibodies under an in-vitro as well as in-cell translation condition. Principally, the present invention is a novel RNA replicase-mediated RNA / mRNA amplification method, namely Replicase Cycling Reaction (RCR). The RNA replicases involved in RCR include but not limited to viral and / or bacteriophage RNA-dependent RNA polymerases (RdRp), particularly coronaviral and hepatitis C viral (HCV) RdRp enzymes.

The invention belongs to the technical field of biology, and particularly discloses a system for detecting HIV RNA, a fluorescent biosensor and preparation and application of the fluorescent biosensor. The system comprises red blood cells, a first report chain, a second report chain, crRNA, Cas12a and a DNA hairpin probe H1. On the basis of the system, a fluid membrane-red blood cell confinement Cas12a can be constructed, and a simple, sensitive and specific fluorescence detection sensing strategy of virus RNA such as HIV RNA is developed, so that the problems of low reaction efficiency, complex reaction steps and the like of an HIV RNA detection method based on CRISPR-Cas12a in the prior art are solved.

The present inventors have conducted intensive studies on an antibody which controls HIV in an administration group with a high probability over a long period of time with one or several times of single-agent administration. As a result, the present inventors have surprisingly found that, when an SW-1C10 antibody, which is obtained by producing an antibody gene reported as 1C10 in silkworms, is singly administered only a few times, the viral load in the blood is suppressed to the detection limit or lower at an early stage in all of individuals to which the antibody has been administered, and moreover, the viral RNA load in the blood is maintained at the detection limit or lower for a long time of 12 weeks.

The invention discloses an influenza reporter virus carrying a fluorescent RNA aptamer and a construction method and application thereof, and the influenza reporter virus carrying the fluorescent RNA aptamer is obtained by introducing a termination codon TAA into the 328th nucleotide of an ORF region of an NS1 gene on the basis of a genome of an influenza A virus and terminating in advance, then introducing a terminating codon TAA into the 328th nucleotide of the ORF region of the NS1 gene of the influenza A virus; original nucleotides at the 328-480 sites are replaced by forward or reverse RNA aptamers, and the RNA aptamer is obtained. The influenza reporter virus carrying the fluorescent RNA aptamer constructed by the invention also has the characteristics of good virus replication ability, high RNA imaging sensitivity and good specificity on the basis of stable inheritance, can be used for real-time visual observation of virus RNA, and is used for research on virus replication cycle and molecular mechanism related to the virus RNA. Besides, the construction method of the influenza reporter virus carrying the fluorescent RNA aptamer can also be applied to construction of RNA fluorescent reporter viruses of other subtype influenza viruses such as H3N2 and H5N1, and an important tool is provided for real-time tracing of corresponding virus RNA.

A protection method, a protection sequence, a composition and a kit for preventing RNA degradation, and the use thereof, which can improve the protective effect on viral RNA. After the protection sequence RNase AP is added, the three-dimensional structure of RNA and the structural allosteric characteristics of RNA can be protected; and the protection sequence has a good protection effect on RNA integrity, and a high-level detection rate can be obtained by a one-time amplification, thereby ensuring efficient enrichment of viral RNA and providing guarantee for subsequent reverse transcription and PCR amplification.

Methods of inhibiting a respiratory virus, (i.e., a virus associated with a respiratory condition, e.g., influenza A, influenza B, RSV, etc.) in a sample are provided. Aspects of the methods include contacting a sample comprising viral RNA (vRNA) having a target motif with an effective amount of an agent that specifically binds the target motif to inhibit the respiratory virus. Also provided are methods of treating or preventing respiratory virus infection in a subject. Also provided are compounds and pharmaceutical compositions comprising an oligonucleotide sequence complementary to a target vRNA region that find use in the subject methods.

The disclosure provides an engineered Salmonella typhimurium comprising a first heterologous nucleic acid encoding a first polypeptide that causes bacterial lysis, a second heterologous nucleic acid encoding a second polypeptide that causes vacuolar lysis, and a third heterologous nucleic acid encoding a virus, wherein the virus comprising a picornavirus. Further disclosed is a method of using the engineered bacteria for treating a tumor in a subject.

The invention relates to an infectious arenavirus particle that is genetically engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but incapable of producing further infectious progeny particles in normal, non-genetically engineered cells.One or more of the four arenavirus open reading frames glycoprotein (GP), nucleoprotein (NP), matrix protein Z, and RNA-dependent RNA polymerase L are removed or mutated to prevent replication in normal cells, but still allow gene expression in arenavirus vector-infected cells, and foreign genes encoding an antigen or other protein of interest or nucleic acids that regulate host gene expression are expressed under the control of arenavirus promoters, internal ribosome entry sites, or under the control of regulatory elements that can be read by the viral RNA-dependent RNA polymerase, cellular RNA polymerase I, RNA polymerase II, or RNA polymerase III. The modified arenaviruses are useful as vaccines and therapeutic agents for a variety of diseases.

The application discloses a kind of RNA virus nucleic acid preservative in fecal sample, including denaturant, chelating agent, pH buffering agent, osmotic pressure regulator and reducing agent;The denaturant includes guanidine salt and surfactant;The application further additionally provides a kind of RNA virus nucleic acid preservative in fecal sample preparation method, preparation above-mentioned one kind of RNA virus nucleic acid preservative in fecal sample, including the following steps: each component is mixed with ratio, pH is adjusted with pH buffering agent, with deionized water constant volume;Filter, obtain the preservative solution described in this application.The preservative solution of the application can stably store human cell DNA and viral RNA in fecal sample at room temperature for at least 7 days, which is beneficial for long-distance transportation at room temperature;Meanwhile, the preservative solution of the application also releases nucleic acid, inactivates virus, so as to prevent biological leakage and infection phenomenon;The de-inhibitor component in the preservative solution can directly combine and precipitate with various inhibitors in feces, reducing the interference to nucleic acid detection.

The invention discloses application of nicotinamide ribose (NR) in preparation of anti-influenza virus products, and belongs to the technical field of biotechnology and medicine. The NR is used as a human endogenous metabolite, has excellent safety, activates SIRT1 by increasing the NAD + level in cells, and further inhibits influenza virus NP protein nucleus input and virus RNA synthesis. Experiments prove that NR has a remarkable inhibition effect on replication of various subtype influenza A viruses such as H1N1, H9N2 and H3N2, still has an inhibition effect under high-MOI infection, has a good use effect at the early stage of infection, and provides a new choice for anti-influenza virus drugs.

The application belongs to the technical field of RNA rapid detection, and particularly relates to a rapid amplification and lateral chromatographic detection method for virus RNA and application. The crude virus nucleic acid extract is subjected to a single-tube composite enzyme reaction system reverse transcription, restriction endonuclease digestion and one-way isothermal strand displacement amplification, and then a large amount of specific sequence single-stranded DNA with a marker is generated; in the lateral chromatographic process, the single-stranded DNA is captured by a nucleic acid probe fixed on a detection area of a virus RNA detection test paper, at the same time, avidin labeled on a colloidal gold particle is combined with biotin labeled on the fixed single-stranded DNA, and the colloidal gold particle is indirectly fixed to show a color indication result. The method provided by the application is simple to operate, does not need complicated nucleic acid extraction, does not need large equipment in the whole detection process, and the result can be judged by naked eyes, so that on-site rapid detection can be realized, and the method has the advantages of simplicity, rapidness, high sensitivity and low cost.

Methods and kits for the rapid detection of active coronavirus infection by specifically targeting the negative-sense RNA produced during viral replication. The method utilizes nucleic acid amplification techniques, such as RT-PCR and qPCR, with primers specific to the negative strand, which serves as a marker for active viral replication. The present disclosure also encompasses a rapid, user-friendly lateral flow test based on CRISPR-Cas13a technology that targets the negative-sense viral RNA. This LFT employs guide RNAs specific to the negative strand and a reporter system to visually indicate the presence of actively replicating viruses, thereby distinguishing active infections from the presence of non-replicating viral remnants. The methods and kits offer a more accurate means of determining infectivity, guiding public health measures, and monitoring antiviral therapies.

The present invention offers a new approach for highly multiplexed, programmable antiviral therapies that directly target viral RNA, and can be flexibly adapted to target novel viruses or emerging outbreak pathogens. Class 2, type VI CRISPR system-based therapies can be used in combination with existing antiviral compounds for viruses where such compounds exist, either by increasing their efficacy or by preventing the evolution of specific drug resistance mutations. Perhaps most excitingly, if a virus evolves resistance to a specific guide RNA sequence, it is easy to switch to a different guide RNA sequence, or to design a new guide sequence to target the new mutation. Such approaches should prevent the widespread development of resistance to Class 2, type VI CRISPR system-based therapies.

The invention discloses lactobacillus mucosa with porcine virus resisting activity and application of the lactobacillus mucosa, and relates to the technical field of microorganism application and virus prevention and control. It is found for the first time that the lactobacillus mucosa has broad-spectrum antiviral activity, not only has a remarkable inhibition effect on porcine reproductive and respiratory syndrome virus type 2 (PRRSV2) strains, but also can further resist porcine epidemic diarrhea virus (PEDV) and porcine rotavirus (PoRV); according to the present invention, the Lactobacillus mucosa is adopted to culture the supernatant, such that the supernatant does not have toxicity on Marc-145 cells; according to the PRRSV2 strain treated by using the supernatant, the expression level of virus RNA and protein and the filial generation virus titer are remarkably reduced, so that the infection ability is weakened; the culture supernatant directly acts with virus particles to block infection and replication of the virus particles; the product disclosed by the invention is derived from natural probiotics, is high in safety, environment-friendly and not easy to resist drugs, can be used for preparing a broad-spectrum anti-porcine virus biological preparation, provides a new strategy for preventing and controlling porcine virus diseases, and has a good industrial application prospect.

A method and composition for the simultaneous detection of a protein and a nucleic acid within a single biological sample using proximity ligation of aptamers and quantitative polymerase chain reaction (qPCR) is disclosed. The invention employs two aptamers, each targeting a distinct epitope of the protein of interest, functionalized with primer sites to enable ligation by way of a short DNA linker in the presence of the protein. This ligated construct is then amplified and detected using qPCR. Concurrently, viral RNA is detected in the same reaction using reverse transcription- qPCR (RT-qPCR) with TaqMan probes in a separate fluorescent channel. This dual detection approach is demonstrated with SARS-CoV-2 nucleocapsid protein and RNA and is validated in saliva samples and digital droplet PCR. The invention provides a sensitive, rapid, and specific method for multiplexed molecular diagnostics.

This invention belongs to the field of biomedical technology, specifically disclosing a scorpion-derived polypeptide for combating influenza virus and its applications. The scorpion-derived polypeptide has the amino acid sequence shown in SEQ ID NO:1, or is a functional variant obtained by substitution, deletion, and / or addition of one or more amino acids from the same sequence, retaining its anti-influenza virus activity. The scorpion-derived polypeptide provided by this invention can significantly inhibit the proliferation of influenza A virus and substantially reduce the expression levels of viral RNA and protein within a non-cytotoxic concentration range. It also effectively reduces viral load and inflammatory factor levels in mice infected with influenza A virus, exhibiting excellent anti-influenza A virus activity and being less likely to induce viral resistance. This invention provides a novel, highly efficient, and low-toxicity bioactive molecule for influenza prevention and control, which can be applied to the development of anti-influenza drugs.

The application provides a phosphoramidate morpholino antisense oligonucleotide for antiviral, which comprises a base sequence shown as SEQ ID NO. 1, the antisense oligonucleotide is a phosphoramidate morpholino antisense oligonucleotide, the structural formula of the phosphoramidate morpholino nucleotide monomer in the antisense oligonucleotide is shown in the following formula, Base is a base; R is N, N-dimethylamine or piperazine. The antisense oligonucleotide can be combined with a specific region in the fifth segment of viral RNA of an influenza virus, block the transcription of the viral RNA through steric hindrance, play an antiviral role, and use a PMO structure which is good in nuclease stability, high in antisense efficiency and good in water solubility, and at the same time, by introducing a piperazine group which is positively charged under physiological conditions, the PMO compound can enter the cell across the membrane, and the biological activity of the compound can be effectively enhanced.

Described herein are methods for the detection of a virus (e.g., SARS-CoV-2) RNA in dust, which can be used for continued environmental surveillance of the viral disease. Targeted monitoring of dust in high-concern buildings can complement broader population-level monitoring approaches. Additionally, a method for detection of a viral RNA in a dust sample is disclosed herein.

The present invention provides methods and kits for detecting and quantifying the presence of porcine virus PERV-A, PERV-B, PCMV, PCV3, PLHV-1, PLHV-2, and PLHV-3 nucleic acids in a sample from a xenotransplant subject that has received cells, tissues or an organ from a pig.

This invention discloses a drug for preventing and treating myocarditis caused by CVB3 virus and its application, belonging to the field of biomedical technology. This invention discovers an hsa-miR-199a-5p mimic that can be used to prevent and treat myocarditis caused by CVB3 virus. The nucleotide sequence of this hsa-miR-199a-5p mimic is shown in SEQ ID NO.1, and its antisense strand nucleotide sequence is shown in SEQ ID NO.2. This invention reveals that hsa-miR-199a-5p mimics can effectively inhibit viral RNA replication and protein biosynthesis and expression in CVB3, and reduce cell death caused by viral infection. Therefore, hsa-miR-199a-5p plays an important role in regulating and controlling CVB3 replication and protein expression. The use of hsa-miR-199a-5p mimics and / or their antisense strands to prepare anti-CVB3 virus drugs, drugs for the prevention and / or treatment of viral myocarditis, drugs for inhibiting CVB3 viral protein expression, drugs for inhibiting CVB3 viral replication, or drugs for reducing CVB3 virus-induced cell death, as well as the preparation of CVB3 virus detection products and viral myocarditis diagnostic products, all have significant clinical application potential.

Type-C endogenous retroviruses (ERVs) embedded in Chinese hamster ovary (CHO) cells were altered to modify the release of retroviral and / or retroviral-like particles in the culture supernatant. Although evidence for the infectivity of these particles is missing. their presence has raised safety concerns. 173 type-C ERV sequences that clustered into functionally conserved groups were identified. Transcripts from one type-C ERV group were identified to be full-length with intact open reading frames, and to have corresponding viral RNA genomes that were loaded into retroviral-like particles. Also, sequence analysis of the genomic RNA from viral particles indicated that they may result from few expressed ERV sequences. Disclosed herein is the disruption / alteration of the gag gene of the expressed ERV group using CRISPR-Cas9 genome editing. Comparison of CRISPR-derived mutations at the DNA and mRNA level led to the identification of a single ERV locus responsible for the release of viral RNA-loaded particles from CHO cells. Clones bearing a Gag loss-of-function mutation in this particular ERV locus showed a reduction of viral RNA-containing particles in the cell supernatant by over 250-fold. Notably, ERV mutagenesis did not compromise cell growth, cell size or recombinant protein production. Provided herein is a new strategy and cells, in particular engineered CHO cells, to mitigate potential contaminations from CHO endogenous retroviruses during biopharmaceutical manufacturing.

Provided are compositions, systems, and methods useful for effecting gene editing in eukaryotic cells. Compositions include plasmids that encode one or more viral fusion proteins in which one or more viral proteins are fused with an aptamer-binding protein. Compositions also include plasmids that encode a non-viral nucleic acid sequence, wherein the non-viral nucleic acid sequence encodes a CRISPR system component. In some instances, the non-viral nucleic acid sequence also includes an aptamer sequence. The plasmids can be used to generate viral particles, including lentivirus-like particles that contain a viral fusion protein and a non-viral RNA sequence. Systems of producing such viral particles are provided. Also provided are methods of using the viral particles of the disclosure to effect gene editing in eukaryotic cells.

The present invention relates to the use of a bacterial artificial chromosome (BAC) for the preparation of a vaccine wherein the BAC comprises:-an inducible bacterial ori sequence for amplifying the BAC to more than 10 copies per bacterial cell, and-a viral expression cassette comprising cDNA of an attenuated RNA viral genome, and comprising a cis-regulatory element for transcribing the viral cDNA in mammalian cells and for processing the transcribed RNA into infectious viral RNA.

The application discloses a chimera specifically targeting and degrading an RNA virus genome and an application thereof, and the chimera is composed of a chimera N and a chimera C; the chimera N is sequentially connected with a MAVS CARD domain, a connecting peptide and an N terminal of RNase A; the chimera C is sequentially connected with a MAVS CARD domain, a connecting peptide and a C terminal of RNase A; and the N terminal of RNase A and the C terminal of RNase A are obtained by segmenting RNase A at a segmentation site. The chimera in the application assembles in situ to form a CARTAC complex with RNA cleavage activity, and then degrades a viral RNA genome, thereby providing a safe, broad-spectrum and effective prevention and treatment strategy for resisting RNA virus infection.

Provided are compositions, systems, and methods useful for effecting gene editing in eukaryotic cells. Compositions include plasmids that encode one or more viral fusion proteins in which one or more viral proteins are fused with an aptamer-binding protein. Compositions also include plasmids that encode a non-viral nucleic acid sequence, wherein the non-viral nucleic acid sequence encodes a CRISPR system component. In some instances, the non-viral nucleic acid sequence also includes an aptamer sequence. The plasmids can be used to generate viral particles, including lentivirus-like particles that contain a viral fusion protein and a non-viral RNA sequence. Systems of producing such viral particles are provided. Also provided are methods of using the viral particles of the disclosure to effect gene editing in eukaryotic cells.

The invention belongs to the technical field of DNA tracks, and particularly relates to a recoverable magnetic DNA track as well as a preparation method and application thereof. Through a magnetic DNA track mediated recognition mode and a label-free detection method in the invention, sustainable detection of virus RNA is realized, the detection limit is as low as 0.9 fM, and high sensitivity and specificity are achieved. Besides, the magnetic DNA track provided by the invention provides an innovative and sustainable platform for detecting virus RNA, and has a great application prospect in the aspects of diagnosis of coronavirus diseases, food safety evaluation, environmental monitoring and the like, and the sustainable DNA track is beneficial to reducing resource waste and economic burden.

The invention relates to an anti-dengue virus medicine containing a ferroptosis inhibitor and application of the anti-dengue virus medicine, in particular to medical application of the ferroptosis inhibitor in resisting dengue virus infection. In-vitro experiments prove that DENV infection can activate a host cell ferroptosis pathway; after 1-3 [mu] M of Ferrostatin-1 or Liproxstatin-1 is adopted for intervention, the activation of the pathway can be effectively inhibited, and the virus RNA replication and protein expression level can be remarkably reduced. According to the technical scheme, the existing activity of the known ferroptosis inhibitor is utilized, new indications for resisting DENV infection of the ferroptosis inhibitor are developed, a candidate drug strategy of a targeted ferroptosis pathway is provided for dengue treatment, the technical problem that the action target of an existing antiviral drug is limited is solved, and the obvious clinical application and development value is achieved.

The present invention relates to a polypeptide composition, comprising a nucleic acid encoding a mutant pp65 polypeptide, the nucleic acid being mRNA, viral RNA, or replicon RNA. The composition is capable of expressing the mutant pp65 polypeptide in vivo, eliciting a specific humoral or cellular immune response against the pp65 polypeptide, and producing an effector cell having a tumor cell killing effect and a specific antibody.

The present invention belongs to the technical field of genetic engineering, in particular to a method for detecting classical swine fever virus based on a G4-ThT biosensor and NASBA and a kit thereof. The specific method includes using the viral RNA extracted from a classical swine fever virus sample as the amplification target, performing isothermal amplification using an isothermal nucleic acid amplification technique, then adding a buffer solution containing Tris-HCl and KCl, and further adding thioflavin T to promote the folding of G-quadruplexes on the amplification products to form a complex, detecting fluorescence under the excitation of a light source with a specific wavelength, and making a judgment according to the intensity thereof. The present invention provides a probe that does not require special biotin labeling or thiol modification treatment, a detection method with significantly reduced detection costs, and a kit.

Methods of inhibiting influenza A virus in a sample are provided. Aspects of the methods include contacting a sample comprising viral RNA (vRNA) having a PSL2 motif with an effective amount of an agent that specifically binds the PSL2 motif to inhibit the influenza A virus. Also provided are methods of treating or preventing influenza A virus infection in a subject. Also provided are methods for screening a candidate agent for the ability to inhibit influenza A virus in a cell, the method comprising: contacting a sample with a candidate agent; and determining whether the candidate agent specifically binds to the PSL2 motif of vRNA. Also provided are compounds and pharmaceutical compositions comprising an oligonucleotide sequence complementary to a PB2 vRNA region that find use in the subject methods.