34 results about "Negative strand" patented technology

The invention relates to an HBV magnetic bead as well as a preparation method and application thereof. The HBV magnetic beads are HBV magnetic beads with molecular tag primer sequences, poly (dT) and N HBV specific capture probes are coupled and modified, and N is a positive integer and is greater than or equal to 1. The HBV specific capture probe is selected from the following groups: (a) a pgRNA probe; (b) an S probe; (c) an X probe; (d) an rcDNA probe; (e) a cccDNA probe; wherein the pgRNA probe targets a positive chain pgRNA region of the HBV, the S probe targets a positive chain S region of the HBV, the X probe targets a positive chain X region of the HBV, the rcDNA probe targets a negative chain rcDNA region of the HBV, and the cccDNA probe targets a negative chain cccDNA region of the HBV. By adopting the novel HBV magnetic bead, the enrichment of HBV sequences can be greatly improved.

The invention belongs to the technical field of biology, and particularly relates to application of carboplatin in preparation of a medicine for preventing and / or treating negative chain RNA virus infection. Experimental results show that the carboplatin has no obvious influence on the cell survival rate, and the virus titer in the Vero cells treated by the carboplatin is obviously reduced; the replication of PPRV in Vero cells can be significantly inhibited, and the inhibition effect is most significant when the concentration of carboplatin is 100 [mu] M; and the N protein level in Vero cells treated by carboplatin (100 [mu] M) is remarkably reduced, so that the method has a wide application prospect.

To provide nucleic acid molecules targeting a BCL11A enhancer functional region, compositions comprising the nucleic acid molecules, and methods for increasing a fetal hemoglobin level in cells by disrupting BCL11A expression at the genomic level.SOLUTION: Disclosed is a nucleic acid molecule comprising a nucleic acid sequence being: a. complementary to the plus or minus strand of the human chromosome 2 at location 60725424 to 60725688 (+55 functional region); b. complementary to the plus or minus strand of the human second chromosome at location 60722238 to 60722466 (+58 functional region); or c. complementary to the plus or minus strand of the human chromosome 2 at location 60718042 to 60718186 (+62 functional region), where the human chromosome 2 is that according to UCSC Genome Browser hg 19 human genome assembly.SELECTED DRAWING: Figure 3G

The present invention provides a temperature-sensitive negative-strand RNA virus or virus vector and an RNA genome thereof. According to the invention, provided is a negative-strand RNA virus or virus vector having a negative-strand RNA genome wherein the phosphoprotein (P protein) on the RNA genome has an amino acid mutation(s) corresponding to a substitution(s) in an amino acid(s) of the P protein corresponding to one or more or all of D433, R434, and K437 and optionally further has an amino acid mutation corresponding to a further amino acid substitution in an amino acid of the P protein corresponding to L511.

The present invention provides a temperature-sensitive negative-strand RNA virus or virus vector and an RNA genome thereof. According to the invention, provided is a negative-strand RNA virus or virus vector having a negative-strand RNA genome wherein the phosphoprotein (P protein) on the RNA genome has an amino acid mutation(s) corresponding to a substitution(s) in an amino acid(s) of the P protein corresponding to one or more or all of D433, R434, and K437 and optionally further has an amino acid mutation corresponding to a further amino acid substitution in an amino acid of the P protein corresponding to L511.

A method for detecting a coronavirus in a sample, the method containing i) isolating RNA from a sample; ii) contacting the isolated RNA with a first primer in the presence of a reverse transcriptase and dNTP under conditions permissive for cDNA synthesis, thereby generating cDNA; iii) contacting the cDNA with forward and reverse primers in the presence of a DNA polymerase and dNTP under condition permissive for cDNA amplification, thereby generating an amplified product, and iv) identifying the amplified product, thereby detecting the presence or absence of the virus, wherein the first primer binds to or hybridizes with a negative-strand RNA molecule produced by the virus.

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.

A method for detecting a coronavirus in a sample, the method containing i) isolating RNA from a sample; ii) contacting the isolated RNA with a first primer in the presence of a reverse transcriptase and dNTP under conditions permissive for cDNA synthesis, thereby generating cDNA; iii) contacting the cDNA with forward and reverse primers in the presence of a DNA polymerase and dNTP under condition permissive for cDNA amplification, thereby generating an amplified product, and iv) identifying the amplified product, thereby detecting the presence or absence of the virus, wherein the first primer binds to or hybridizes with a negative-strand RNA molecule produced by the virus.

The invention relates to the technical field of medicines, in particular to application of DNMT3L gene coded protein in preparation of antiviral drugs, the DNMT3L coding gene is proved to have the effect of inhibiting replication and transcription of linear double-stranded DNA viruses or single-stranded negative-stranded RNA viruses, and analysis shows that cells over-expressing the DNMT3L coding gene can promote interferon expression, so that the anti-virus effect of the DNMT3L coding gene is improved, and the anti-virus effect of the DNMT3L coding gene is improved. According to the present invention, the DNMT3L coding gene is constructed into the overexpression plasmid by using the molecular biology method so as to significantly inhibit the replication of the linear double-chain DNA virus or the single-strand negative-chain RNA virus, and the DNMT3L coding gene is overexpressed in the host cell by using the vector so as to enhance the cell antiviral ability;

The application discloses a detection method based on dominant clones caused by exogenous DNA insertion mutation, and the method comprises the following steps: 1, preprocessing of virus IS information data by positive and negative strand correction and redundant integration; 2, after the preprocessing is completed, IS caused by non-specific amplification is identified to remove false positive IS detection results caused by PCR non-specific amplification; 3, according to a site clustering algorithm, IS in adjacent positions is combined and clustered; 4, according to the clustering results, a clone plane is analyzed, and dominant clones are described; and the application utilizes a PCR primer similarity ranking algorithm to identify and remove non-specific amplification caused by possible PCR primer similarity, so that the false positive of IS identification is significantly reduced in actual application, and the accuracy is improved.

The present invention belongs to the technical field of antiviral drugs, and particularly relates to the application of astaxanthin in the preparation of antiviral products. The viruses include DNA viruses, positive single-stranded RNA viruses, and single-stranded negative-strand RNA viruses. Through in vitro and in vivo experiments, the present invention has confirmed that astaxanthin can inhibit virus adsorption and internalization, reduce the damage of the virus to the cell membrane structure and function, astaxanthin can inhibit virus invasion, and directly reduce the virus titer in animals. The research results of the present invention expand the scope of the efficacy of astaxanthin, and for the first time confirm that astaxanthin can prevent and treat virus-induced infections and diseases.

The invention relates to the field of protein, in particular to a protein composition for detecting ADAMTS13 activity and application of the protein composition. The protein combination for detecting the activity of the ADAMTS13 is composed of two protein subchains, namely, Chain A and Chain B. The Chain A comprises annularly arranged GFP as shown in SEQ ID NO.1, VPart1 of a vWF variant as shown in SEQ ID NO.2 and negative chain polypeptide as shown in SEQ ID NO.5, and the Chain B comprises GFP as shown in SEQ ID NO.1, VPart1 of the vWF variant as shown in SEQ ID NO.2 and negative chain polypeptide as shown in SEQ ID NO.5. The Chain B comprises a nano antibody as shown in SEQ ID NO.6, a V Part2 of a vWF (D1596-R1668) variant as shown in SEQ ID NO.3, and a positive chain polypeptide as shown in SEQ ID NO.4. The invention also discloses a preparation method of the chain B. The protein combination for detecting the activity of the ADAMTS13 can be used for directly determining the activity of the ADAMTS13 enzyme in a blood sample, a biological sample or a chemical sample, and is suitable for clinical instant inspection, biological sample screening and drug research and development scenes.

A method, device, medium and program for detecting mutations on the basis of methylation data. The method comprises obtaining methylation data of a sample to be tested; on the basis of the methylation data, obtaining base composition data of a 3-base genome after chemical or enzymatic conversion; on the basis of the base composition data of the 3-base genome after chemical or enzymatic conversion, determining base composition data of a 4-base genome before chemical or enzymatic conversion according to the complementary base-pairing relationship between a positive strand and a negative strand, and / or the corresponding relationship between genotypes before and after chemical or enzymatic conversion; on the basis of the base composition data of the 4-base genome, identifying candidate mutation sites; and filtering the candidate mutation sites to generate detection results relating to single nucleotide variation mutation sites. According to the method, the single nucleotide variations can be detected from whole genome methylation data, and the accuracy of detection for low abundance mutations can be significantly improved.

The application discloses a Zika virus inhibiting miRNA-A and application thereof, and belongs to the technical field of biology. The sequence of the miRNA-A is as follows: a positive strand: 5'-UUAGUGUUGUCAGGCCUGCUA-3'; and a negative strand: 5'-UAGCAGGCCUGACAACACUAA-3'. Compared with the known vsiRNA-1 sequence in the prior art, the vsiRNA-A provided by the application has a better Zika virus inhibiting effect, and more choices are provided for the treatment of Zika virus related diseases.

The invention relates to the field of vaccine research and development, in particular to construction of an Ebola virus recombinant vaccine based on a recombinant measles virus vector, a reverse genetic system and application of the Ebola virus recombinant vaccine. Specifically, a measles virus is used as a vector, and a recombinant vaccine of an Ebola virus related antigen is constructed and expressed through a reverse genetic technology. The vaccine can be used for preventing Ebola virus infection and has a wide application prospect. In addition, the invention also discloses a recombinant measles virus vector, a nucleic acid molecule, a reverse genetic system, a recombinant measles virus particle and application thereof. The measles virus vector is characterized in that a negative strand RNA coding sequence of Ebola virus glycoproteins GP and VP40 is inserted into the measles virus vector. The measles virus vector-based Ebola virus vaccine provided by the invention has high immunogenicity and broad spectrum, and a new idea is provided for the development of novel Ebola virus vaccines. The vaccine provided by the invention is simple in preparation method and can be produced on a large scale in a short time.

The invention belongs to the technical field of gene engineering, and relates to a gRNA library of a targeted SARS-CoV-2 virus subgenome. The library is obtained through multi-step rational design screening, is composed of 65 gRNAs, can specifically recognize and target positive chain and negative chain subgenome RNAs of viruses ORF3a, E, M, ORF6, ORF7a, ORF7b, ORF8, N and ORF10, and has a sequence as shown in SEQ ID NO: 1 to 65. The gRNA can form a CRISPR-Cas13b (clustered regularly interspaced short palindromic repeats-associated protein 13b) system with a Cas13b protein, and is used for non-therapeutically inhibiting the replication of the SARS-CoV-2 virus.

The invention belongs to the technical field of virus detection, and particularly relates to a method for marking a positive single-stranded RNA (Ribonucleic Acid) virus replication site in a plant living cell, which comprises the following steps: by taking pAB1677 plasmid as a template, amplifying dCas13BT3 by utilizing primers shown in SEQ ID NO.1 and SEQ ID NO.2, and connecting the dCas13BT3 into a fluorescent protein expression vector to obtain a recombinant vector; preparing gRNA of a target positive single-stranded RNA virus negative strand; and co-transferring the recombinant vector, the gRNA targeting the negative strand of the positive single-stranded RNA virus and the RNA of the positive single-stranded RNA virus into a plant, culturing, and observing the fluorescent protein expression position which is the replication site of the positive single-stranded RNA virus. According to the present invention, by designing the gRNA targeting the PnMV negative chain RNA and using the fusion of dCas13BT3 and the fluorescent protein, the fluorescence signal is accurately enriched in the virus replication complex, such that the background noise problem caused by the positive chain RNA targeting is effectively overcome;

The application discloses a library sequencing method for determining methylation difference of positive and negative strands of DNA molecules. The application provides a library sequencing method for determining methylation difference of positive and negative strands of DNA molecules, comprising the following steps: 1) using molecular tags UMI to label positive and negative strands of a plurality of DNA double-stranded molecules to obtain a DNA molecule library labeled with molecular tags; 2) performing bisulfite conversion on DNA molecules in the DNA molecule library, converting unmethylated modified C bases in the DNA molecules into U bases to obtain converted DNA molecules; 3) sequentially performing PCR amplification, constructing a sequencing library and sequencing on the converted DNA molecules; and determining the difference in methylation modification of positive and negative strands of the same DNA molecule. Compared with traditional whole genome methylation sequencing, the application can more accurately detect the methylation difference of DNA molecules, and more accurately detect the dynamic change of methylation.

The embodiment of the invention relates to a method, a device, equipment and a product for determining the miss probability of a probe sequence. The method includes constructing a first genome comprising at least one of a low methylation positive strand, a low methylation negative strand, a high methylation positive strand, and a high methylation negative strand. The method also includes determining an off-target probability of the probe sequence based on the constructed first genome. Through the method, the accuracy and sensitivity of off-target detection of the probe sequence are improved, false matching or missing detection is effectively avoided, and a basis is provided for specific control of probe sequence design.

Macrophages (Mφ) are crucial for pathogen detection and elimination, and also serve as a microenvironment for replication of multiple infectious pathogens. This delicate balance between viral replication and antiviral response remains largely unknown at the molecular level. Addressing this issue is physiologically important given the persistent threat posed by emerging viral pathogens. Here, we demonstrate that GAS7 expression within macrophages restricts the replication of viral pathogens belonging to all major viral groups. We show that the antiviral activity of GAS7 exists even under conditions where the classical antiviral response mediated by type I interferon is neutralized. In particular, we show that in human monocyte-derived macrophages, silencing of GAS7 boosts the replication of multiple viral pathogens representing most relevant viral groups. These include the retroviruses HIV-1 (human immunodeficiency virus type 1) and HIV-2 (human immunodeficiency virus type 2), the RNA viruses Zika virus (positive-sense single-stranded RNA), Sindbis virus (single-stranded positive-sense RNA), Sendai virus (negative-sense single-stranded RNA), VSV virus (negative-sense single-stranded RNA), and measles virus (negative-sense single-stranded RNA), and the DNA virus HSV-1 virus (double-stranded DNA). Importantly, the antiviral activity of GAS7 is present even under conditions where type I interferon is neutralized by the addition of the B18R protein (which efficiently blocks the type I receptor). Furthermore, the inventors demonstrate that by forcing macrophages to overexpress GAS7, they are further protected from HIV-1 infection compared to cells expressing this factor at normal levels. Therefore, the present invention relates to the modulation of GAS7 expression and / or activity for modulating viral replication in macrophage populations.

The invention provides an integrated DNA interference excluding in-situ hybridization probe and an in-situ hybridization method for detecting hepatitis B virus cccDNA, the in-situ hybridization probe is designed based on a Basescope detection method, and comprises an HBV cccDNA probe specifically targeting an integrated breakpoint of HBV negative chain DNA, an HBV DNA probe targeting an HBV negative chain, an HBV cccDNA probe targeting an HBV negative chain, an HBV cccDNA probe targeting an HBV negative chain, an HBV cccDNA probe targeting an HBV negative chain, and an HBV cccDNA probe targeting an HBV negative chain. The HBV RNA probe with the targeted HBV positive strand gap structure and the matched ribonuclease of the HBV RNA probe can be used for detecting cccDNA and integrating HBV DNA after being treated, and the targeting sequences of the HBV cccDNA, the HBV DNA and the HBV RNA probe are respectively shown as SEQ ID No.1 to SEQ ID No.3. The in-situ hybridization probe and the in-situ hybridization method can be applied to screening medicines for preventing or treating hepatitis B virus infection and researching the life cycle of the hepatitis B virus, so that the understanding on the characteristics of the hepatitis B virus is promoted, and the evaluation on a treatment strategy is optimized.

The invention discloses miRNA-A for inhibiting Zika virus and application of the miRNA-A, and belongs to the technical field of biology. The sequence of the miRNA-A is as follows: a positive chain is 5 '-UUAGUGUUGUCAGGCCUGCUA-3', and a negative chain is 5 '-UAGCAGGCCUGCACAACACA-3'. Compared with the known vsiRNA-1 sequence in the prior art, the vsiRNA-A provided by the invention has a better effect of inhibiting the Zika virus, and provides more choices for the treatment of diseases related to the Zika virus.

The present application relates to the field of proteins, in particular to a protein combination for detecting ADAMTS13 activity and application thereof.The protein combination for detecting ADAMTS13 activity provided by the present application is composed of two protein subchains Chain A and Chain B, wherein Chain A comprises a circularly arranged GFP as shown in SEQ ID NO.1, a V Part1 of a vWF variant as shown in SEQ ID NO.2 and a negative chain polypeptide as shown in SEQ ID NO.5; Chain B comprises a nanobody as shown in SEQ ID NO.6, a V Part2 of a vWF (D1596-R1668) variant as shown in SEQ ID NO.3 and a positive chain polypeptide as shown in SEQ ID NO.4.The protein combination for detecting ADAMTS13 activity provided by the present application can directly determine the ADAMTS13 enzyme activity in a blood sample, a biological sample or a chemical sample, and is suitable for clinical point-of-care testing, biological sample screening and drug development scenarios.

To provide a method for producing negative-strand RNA viral vector.SOLUTION: The present invention provides a method for producing a negative-strand RNA virus or viral vector, comprising: expressing a factor from a gene encoding a protein kinase R (PKR)-inhibitory factor operably linked to a control sequence, and supplying the factor to a packaging cell; allowing the packaging cell to express the genomic RNA of the negative-strand RNA virus or viral vector, thereby forming a negative-strand RNA virus or viral vector in the presence of the factor; and recovering the formed negative-strand RNA virus or viral vector; the PKR-inhibitory factor being a PKR-inhibitory viral factor, or nc886 or p58IPK, and the relationship between the virus or viral vector and the PKR-inhibitory factor being heterologous, and / or the relationship between the control sequence and the PKR-inhibitory factor being heterologous.SELECTED DRAWING: None

To develop novel therapeutic techniques that do not pose the risk of genotoxicity to help infertility patients. [Solution] A single-stranded negative-strand RNA viral vector containing a nucleic acid sequence encoding a protein associated with the formation or development of germ cells, or a pharmaceutical composition for preventing or treating infertility containing an RNA molecule encoding the protein, is provided.

The present invention relates to a recombinant replication-deficient or replication-competent negative-strand RNA virus that can (i) present a heterologous antigen on its surface and induce mucosal immunity in a host, (ii) produce a heterologous antigen after invading host cells, thereby improving the host's immune response to the antigen, and (iii) have reduced anti-vector antigenicity compared to a wild-type vector, thereby being able to reduce the host's immune response to the vector.

To provide a temperature-sensitive negative-strand RNA virus or a viral vector, and an RNA genome thereof.SOLUTION: A negative-strand RNA virus or viral vector is provided, in which at least one viral protein encoded in the genome (preferably the P protein or a temperature-sensitive P protein) is in the form of a fusion protein with a BromoTag.SELECTED DRAWING: None

This invention discloses a Zika virus-inhibiting siRNA-B and its applications, belonging to the field of biotechnology. The sequence of the siRNA-B is as follows: positive strand: 5'-UGUGGUGAAAUCCAUGGUUUC-3'; negative strand: 5'-GAAACCAUGGAUUUCACCACA-3'. Compared with the known vsiRNA-1 sequence in the prior art, the vsiRNA-B provided by this invention has a better inhibitory effect on Zika virus, providing a more effective drug for the treatment of Zika virus-related diseases.

The present invention provides temperature sensitive negative strand RNA viruses or viral vectors and RNA genomes thereof. According to the present invention, a negative-strand RNA virus or a viral vector can be provided in which at least one of the viral proteins encoded by the genome (preferably the aforementioned P protein or temperature-sensitive P protein) is in the form of a fusion protein with a bromine tag.

The invention relates to the field of proteins, in particular to a protein for detecting ADAMTS13 activity and application of the protein. The protein for detecting the activity of the ADAMTS13 at least comprises A1) a nano antibody as shown in SEQ ID NO. 1; a) is a vWF (D1596-R1668) variant as shown in SEQ ID NO.2, A3) is a positive chain polypeptide as shown in SEQ ID NO.3, A4) is a negative chain polypeptide as shown in SEQ ID NO.4, and A5) is annularly arranged GFP (cpGFP) as shown in SEQ ID NO.6. The protein for detecting the activity of ADAMTS13 can directly measure the activity of ADAMTS13 enzyme in a blood sample, a biological sample or a chemical sample, and is suitable for clinical point-of-care testing (POCT), biological sample screening and drug research and development scenes.