76 results about "Nucleoprotein" patented technology

The invention belongs to the field of biological prevention and control, and discloses an insect specific virus NLBV3 and application thereof in reducing transmission of rice odontodwarf virus of brown planthopper. The NLBV3 virus genome comprises three chains, the L chain encodes RNA dependent RNA polymerase (RdRP), the M chain encodes glycoprotein precursor (GP), the S chain encodes nucleoprotein (NP), and the nucleotide sequences are respectively shown as SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3; the application is realized by releasing the NLBV3-carrying brown planthopper population, and according to the screening and detection method of the NLBV3-carrying brown planthopper high-virus population provided by the invention, natural brown planthoppers can be effectively infected after the NLBV3-carrying brown planthopper high-virus population is released, the RRSV transmission capability of the brown planthoppers is reduced, and the virus is prevented from damaging rice production. The NLBV3 provided by the invention is an insect specific virus, is harmless to mammals, fishes and shrimps, natural enemy insects and pollination insects, and has the advantages of specific action object, environment friendliness and the like.

The invention discloses application of fluorinated ionizable lipid as a gene drug delivery carrier, and belongs to the technical field of biological medicine. The gene drug delivery carrier provided by the invention is prepared from fluorinated ionizable lipids, cationic lipids, auxiliary lipids, structural lipids and PEG lipids, wherein the molar percentage of the fluorinated ionizable lipids in the gene drug delivery carrier is 1%-10%. According to the invention, the molar percentage of the fluorinated ionizable lipid in the total lipid is optimized to 1-10%, so that the cytotoxicity is obviously reduced, and more excellent nucleic acid transfection efficiency is realized in vivo and in vitro; various nucleic acid drugs including DNA, mRNA, siRNA and CRISPR-Cas9 ribonucleoprotein can be efficiently delivered, action targets of the nucleic acid drugs are expanded to cell nucleuses and cytoplasm, and the nucleic acid drugs are more accordant with the action mechanism of modern gene therapy and editing.

The invention provides a method for cultivating anti-aging glutinous highland barley, highland barley starch as well as a preparation method and application of the highland barley starch, and particularly relates to a method for improving the starch quality character of an existing high-quality glutinous highland barley variety, in particular to a method for cultivating anti-aging glutinous highland barley by utilizing a gene editing technology without exogenous DNA (Deoxyribose Nucleic Acid) integration. A method for fixed-point knockout of an SSIIa gene to cultivate an anti-aging glutinous highland barley new strain under the background of a "Zhengnongglutinous 232" variety comprises the following steps: providing an acceptor material derived from a highland barley variety "Zhengnongglutinous 232" with a glutinous character, and deleting a Wx gene function of the "Zhengnongglutinous 232"; the method comprises the following steps: assembling guide RNA (Ribonucleic Acid) designed aiming at an SSIIa gene and Cas9 protein in vitro to form a ribonucleoprotein RNP complex; introducing the RNP complex into cells of an acceptor material so as to enable Cas9 protein to generate double-chain breakage at a target site of the SSIIa gene; carrying out culture and plant regeneration on the introduced cells to obtain a T0-generation plant of which the SSIIa gene is edited; and screening plants with the function loss of the SSIIa gene, wherein the plants keep the glutinous character of adult agricultural glutinous 232.

The invention relates to lipid nanoparticles for fish in-vivo skin gene editing and application of the lipid nanoparticles, and belongs to the technical field of fish gene editing. The lipid nanoparticle for fish in-vivo skin gene editing comprises a gene editing element and a lipid layer, wherein the lipid layer wraps the surface of the gene editing element; the gene editing element is a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 (CRISPR associated protein 9) ribonucleoprotein compound or nucleic acid for coding the compound; the lipid nanoparticle carrier comprises ionizable cationic lipid, auxiliary lipid, cholesterol, pegylated lipid and permanent cationic lipid, and the lipid nanoparticle carrier comprises the following components in parts by mole: 15 parts of ionizable cationic lipid, 25-30 parts of auxiliary lipid, 30 parts of cholesterol, 3 parts of pegylated lipid and 7 parts of permanent cationic lipid. The formula is high in pertinence, high in editing efficiency, simple, convenient and safe to operate and wide in application prospect.

The application discloses a specific nucleic acid aptamer for a heterogeneous core ribonucleoprotein hnRNPA2B1 and application, and belongs to the technical field of nucleic acid aptamer. The hnRNPA2B1 aptamer is a small single-stranded DNA screened from a designed oligonucleotide library through an in-vitro exponential enrichment ligand system evolution (SELEX), is composed of 80 nucleotides, and has high affinity to the target. The hnRNPA2B1 aptamer can be applied in a detection method or a detection product of the hnRNPA2B1 protein, and a targeted regulation drug or a biological preparation, and can be specifically made into a molecular probe, used as a detection reagent, used as a targeted preparation and the like for diagnosis and treatment of diseases related to the hnRNPA2B1 protein. The hnRNPA2B1 aptamer has strong affinity and specificity, has the advantages of low production cost, stability, convenient synthesis and easy modification compared with an antibody, and is a potential substitute for the antibody in disease diagnosis and treatment.

This invention discloses a gene editing method and application for *Trichoderma reesei* based on a ribonucleoprotein complex. It aims to overcome the problems of low editing efficiency, easy generation of heterozygotes, and risk of exogenous DNA integration in *Trichoderma reesei* using plasmid-based CRISPR / Cas9 systems. This invention combines a dual-site cleavage strategy guided by double sgRNAs with direct RNP delivery. The method involves: designing two guide RNAs spaced 50-700 bp apart for the target gene, pre-assembling them with Cas9 protein in vitro to form an active RNP complex; delivering this complex along with exogenous donor DNA to *Trichoderma reesei* protoplasts. The exogenous donor DNA contains a left homologous arm, an editing sequence, and a right homologous arm, wherein the left and right homologous arms are homologous to the genomic sequences outside the dual cleavage sites on the target gene determined by the double sgRNAs, respectively; upon entering the cell, the RNP complex introduces dual-site cleavage at the target site, achieving precise editing under the guidance of the exogenous donor DNA.

The application provides a double-stranded walking type DNA nanomachine and a preparation method and application thereof, and relates to the technical field of fluorescent analysis and detection.The double-stranded walking type DNA nanomachine for ctDNA fluorescent detection comprises a gold nanoparticle carrier, an anchor probe, a FAM fluorescent reporter probe, a double-stranded walking arm, a Cas12a protein and a crRNA to form a ribonucleoprotein complex.The walking arm of a conventional CRISPR / Cas12a nanomachine is a DNA single-stranded structure, which is easily cut in trans by activated Cas12a, thereby reducing the stability and sensitivity of the nanomachine.The application introduces a double-stranded DNA structure into the walking arm design to replace the conventional DNA single-stranded walking arm, constructs a double-stranded walking type three-dimensional DNA CRISPR / Cas12a nanomachine, effectively resists the reverse cutting of Cas12a on the walking arm, maintains the structural integrity and functional stability of the nanomachine, and greatly improves the detection sensitivity and specificity of lung cancer ctDNA, thereby having a good clinical application prospect.

The application provides an antisense oligonucleotide Jag-i9 ASO and application thereof, relates to the technical field of biological medicine, and the antisense oligonucleotide Jag-i9 ASO has the sequence of 5'-ACTGGGCCCTGCACCTGA-3'. By providing the antisense oligonucleotide of a specific sequence, the binding site of target heterogeneous ribonucleoprotein K and the Jag2 gene is targeted, the expression of the pro-inflammatory Jag2 subtype is inhibited, the Jag2 gene splicing site can be accurately targeted, the heterogeneous ribonucleoprotein K binding function is specifically blocked, a targeted intervention means is provided for the treatment of myocardial ischemia-reperfusion injury, the generation of the pro-inflammatory subtype is inhibited by efficiently and accurately regulating the alternative splicing of a specific gene, myocardial cell apoptosis and inflammatory response are reduced, myocardial function is improved, and a new approach is provided for the prevention and treatment of myocardial ischemia-reperfusion injury, the treatment effect is improved, and the advantages of reducing side effects are achieved.

This invention relates to lipid nanoparticles for in vivo skin gene editing in fish and their applications, belonging to the field of fish gene editing technology. The lipid nanoparticles for in vivo skin gene editing in fish comprise: a gene editing element and a lipid layer, wherein the lipid layer coats the surface of the gene editing element; the gene editing element is a CRISPR / Cas9 ribonucleoprotein complex or a nucleic acid encoding the complex; the lipid nanoparticle carrier comprises ionizable cationic lipids, auxiliary lipids, cholesterol, polyethylene glycol-modified lipids, and permanently cationic lipids, and by molar weight, the proportions are 15 parts ionizable cationic lipids, 25-30 parts auxiliary lipids, 30 parts cholesterol, 3 parts polyethylene glycol-modified lipids, and 7 parts permanently cationic lipids. The formulation of this invention is highly targeted, has high editing efficiency, is simple and safe to operate, and has broad application prospects.

A solution has been discovered that provides a more effective Coronavirus vaccine. The solution is an mRNA vaccine encoding a SARS-CoV-2 nucleoprotein (N) (mRNA-N) in combination with an mRNA vaccine encoding SARS-CoV-2 spike protein(S) (mRNA-S). Chemically modified mRNA-N (pseudouridine) and / or chemically modified mRNA-S (pseudouridine) can be synthesized and packaged in lipid nanoparticles (LNP). In mouse and hamster models, it was shown that mRNA-N alone is immunogenic and can significantly diminish viral loads in the mouse lung after prime-boost intramuscular immunization. In addition, the combinatorial mRNA-N / mRNA-S vaccination induces substantially stronger protection against SARS-CoV-2 than vaccination with mRNA-S alone.

The present disclosure provides RNA-guided CRISPR-Cas effector proteins, nucleic acids encoding same, and compositions comprising same. The present disclosure provides ribonucleoprotein complexes comprising: an RNA-guided CRISPR-Cas effector protein of the present disclosure; and a guide RNA. The present disclosure provides methods of modifying a target nucleic acid, using an RNA-guided CRISPR-Cas effector protein of the present disclosure and a guide RNA.

This invention belongs to the fields of microbial technology and veterinary pharmacology, and discloses a strain of *Alternaria alternata* isolated from Artemisia argyi, its fermentation supernatant, and its application in the preparation of anti-PRRSV drugs. The strain's preservation number is CCTCC NO: M2026443. The fermentation supernatant of this strain can inhibit PRRSV infection of cells and inhibit viral proliferation on cells. Cell-level anti-infection studies show that this strain has significant anti-PRRSV activity in vitro, reducing viral titer, viral genome copy number, and viral nucleoprotein production. It can be used to prepare a broad-spectrum anti-porcine reproductive and respiratory syndrome virus drug to control the occurrence of porcine reproductive and respiratory syndrome.

The invention relates to a ribonucleoprotein complex comprising a nuclease characterized as CRISPR-associated protein of the type II-B having a sequence identity of at least 70% to SEQ ID NO: 1, and a dual guide RNA comprising a CRISPR RNA and a transactivating crRNA, or single guide RNA having a sequence identity of at least 70% to SEQ ID NO: 5. Likewise, the present invention also relates to the use of said ribonucleoprotein to bind, modify or alter target sequences in the genome of a 0 cell or organism, or to modify the gene expression of a cell.

The present invention relates to a DNA aptamer specifically binding to an alpha-synuclein protein and uses thereof. Specifically, the present invention relates to a DNA aptamer selected from the group consisting of nucleotide sequences of SEQ ID NOS: 1 to 12 and binding specifically to an α-synuclein protein, and a composition for detecting an α-synuclein protein, a detection kit, a detection chip or a microarray, all comprising the aptamer as an active ingredient. In addition, the present invention relates to a method for detecting an α-synuclein protein, a method for providing information for diagnosing a degenerative brain disease, a composition for diagnosing a degenerative brain disease, and a pharmaceutical composition for preventing or treating a neurodegenerative disease, all using the DNA aptamer of the present invention.

The invention relates to two kinds of siRNA (small interfering Ribonucleic Acid) targeting heterogeneous ribonucleoprotein C, wherein the siRNA comprises si-Hnrnpc-1 and si-Hnrnpc-2. Compared with the prior art, the siRNA disclosed by the invention has the advantages that the mRNA accessibility, space occupying factors and gene functional regions are considered, the probability of miss of family and cross-family miss can be reduced, Hnrnpc expression can be effectively reduced, meanwhile, the siRNA has clear targeting and biological effects, and the siRNA has important research value and application prospect for future cancer mechanisms and anti-cancer treatment.

The invention provides a measles virus nucleoprotein specific nano antibody as well as a preparation method and application thereof. The amino acid sequence of the nano antibody is any one of A1-A18; the rash virus nucleoprotein can be specifically recognized and combined. And the nano antibody can be massively expressed and purified in escherichia coli cells, so that a potential nano antibody drug with a great application prospect is provided for clinical prevention, treatment and detection of measles viruses.

Provided herein are SNCA antisense oligonucleotides (ASOs) and SNCA ASO conjugates for modulating expression of a SNCA target nucleic acid. Also provided herein are methods of use thereof.

This invention discloses the application of an endogenous Bunyavirus element LsBuEVE and its dsRNA from the planthopper in blocking the transmission of rice viruses. The LsBuEVE genome contains the nucleotide sequence shown in SEQ ID NO.1, and its corresponding transcript contains a complete open reading frame, the nucleotide sequence of which is shown in SEQ ID NO.2, and the encoded amino acid sequence is shown in SEQ ID NO.3. Introducing the dsRNA targeting LsBuEVE into the planthopper effectively inhibits LsBuEVE expression, significantly reduces the transcriptional level of the RSV nucleoprotein gene, protein accumulation, and the RSV acquisition rate in the planthopper, thereby effectively blocking RSV replication and transmission within the planthopper. This invention reveals for the first time the important role of the endogenous viral element LsBuEVE in the transmission of plant viruses by vector insects and provides a precise targeted control strategy based on RNA interference. Compared with traditional chemical pesticides, this technology has advantages such as high specificity, environmental friendliness, and low likelihood of developing resistance, and has broad application prospects and commercial value.

The present invention relates to the use of small artificial RNA (hereafter referred to as "smartRNA") containing sequences creating secondary structure motifs similar to C / D box small nucleolar RNAs (snoRNAs), and antisense guide sequences, as a tool for targeted upregulation of protein expression. The smartRNAs assembles into a ribonucleoprotein (RNP) complex resulting in a smartRNP, increasing the 2´-O-methylation level of one or more adenosines of the polyA-tail of the target mRNA, thereby delaying deadenylation of the targeted mRNA, resulting in increased protein production.

A universal influenza mRNA vaccine is provided, the protein encoded by the universal influenza mRNA vaccine consists of the extracellular domain (M2e), hemagglutinin (HA) stem LAH region, and nucleoprotein (NP) of the influenza A virus matrix protein 2. Furthermore, the application of the universal influenza mRNA vaccine in a mouse animal model is further provided, the universal influenza mRNA vaccine can induce a robust humoral and cellular immune response, protecting mice in the animal model from infection by multiple influenza A viruses. The universal influenza mRNA vaccine is safe, effective, and has a rapid production platform.

The invention belongs to the technical field of biological breeding, and particularly relates to a vegetable seed seedling cultivation method based on gene editing, which comprises the following steps: constructing a targeting nano delivery carrier loaded with gene editing ribonucleoprotein (RNP), the carrier is composed of surface modified polyethyleneimine and carbon nanodots of stem tip meristem homing peptide (MHP), and the carbon nanodots are uniformly distributed on the surface modified polyethyleneimine and the stem tip meristem homing peptide (MHP). RNP is delivered to stem tip meristem of a vegetable seedling through a micro-droplet in-situ permeation method, so that instantaneous editing of a target gene is realized, and homozygous editing seeds without exogenous gene residues are obtained through sexual reproduction. By means of the scheme, non-transgenic gene editing without genetic transformation and tissue culture is achieved, and the method has the advantages of being easy and convenient to operate, short in period, high in efficiency and wide in applicability and is suitable for character improvement of various vegetables such as tomatoes, spinach and cucumbers.

Provided herein are engineered CasX systems and components thereof, including variant CasX proteins and variant guide nucleic acids (gNAs). The variant CasX proteins and variant gNAs of the present invention exhibit at least one improved characteristic when compared to a reference CasX protein or reference gNA of the invention. In some cases, the variants have one or more improved CasX ribonucleoprotein complex functions. Methods of making and using the variants are also provided.

The present invention relates to a method for rescue of Vesicular Stomatitis Virus (VSV) from DNA in a HEK293 cell line or a HEK293 cell line adapted to suspension growth comprising (a) providing cells from a HEK293 cell line or a HEK293 cell line adapted to suspension growth in cell culture, (b) transfecting the cells with at least one plasmid, wherein the at least one plasmid comprises (i) an expression cassette comprising a VSV genomic cDNA; (ii) at least one expression cassette encoding VSV nucleoprotein (N) protein, VSV phosphoprotein (P) protein, and VSV large (L) protein; and (iii) an expression cassette encoding SV40 Large T antigen; (c) culturing the transfected cells; and (d) harvesting the cell culture supernatant comprising the rescued VSV. Also provided is the use of a HEK293 cell line or a HEK293 cell line adapted to suspension growth for rescue of Vesicular Stomatitis Virus (VSV) or the use of a plasmid encoding SV40 Large T antigen for rescue of Vesicular Stomatitis Virus (VSV) in a HEK293 cell line or a HEK293 cell line adapted to suspension growth HEK293-F cells by means of transient transfection.

A method for inserting a polynucleotide exogenous transgene sequence at a predetermined endogenous genetic locus in the genome of a host cell, the method comprising: (i) a donor DNA template comprising a polynucleotide insertion fragment; 5'homologous arms; and a 3'homologous arm. In some embodiments, the 5'homologous arm and the 3 'homologous arm are complementary to DNA in the target region; and (ii) a ribonucleoprotein complex (RNP) comprising (1) a Cas nuclease and at least one small guide RNA (sgRNA) complementary to at least one selected nucleic acid sequence within the predetermined genetic locus in the host cell genome.

The invention discloses an AI auxiliary remote consultation system for testicular nucleoprotein carcinoma, and relates to the technical field of medical information, and the system comprises a multi-modal data fusion processing module which is used for receiving and preprocessing patient multi-modal medical data from different medical institutions; the dynamic feature extraction and tumor evolution modeling module is used for extracting dynamic pathological features related to the testicular nucleoprotein cancer from the fused multi-dimensional feature tensor; the core of the expert diagnosis logic deep simulation module is a deep reinforcement learning network which is pre-trained and continuously learns online; and the real-time interaction and decision synchronization module is used for coordinating data streams and control signals among the modules in the system and managing real-time communication with a remote terminal user. The system solves the problems that an existing system is insufficient in testicular nucleoprotein cancer multi-modal data fusion capability, poor in rare case generalization and low in real-time decision-making efficiency, and the diagnosis accuracy and the remote cooperation efficiency are improved.

The application belongs to the technical field of biological medicine, and provides application of an anti-Tachyirus 1 nucleoprotein inhibitor, and proposes application of a compound candesartan as an anti-Tachyirus 1 nucleoprotein inhibitor. The compound candesartan is obtained in the following manner: through expression and purification of Tachyirus 1 nucleoprotein by using an E. coli system, and through screening of natural medicines and FDA medicine libraries by using a molecular thermal shift assay (TSA) technology, an effective screening system is established, and the compound candesartan capable of interacting with the nucleoprotein is discovered; further, a biological membrane interference technology (BLI) and other experiments are used to determine the affinity, analyze the affinity of the compound candesartan and the nucleoprotein, and simultaneously verify and analyze the function characteristics of the nucleoprotein, so as to verify that the compound candesartan can effectively inhibit the combination of the nucleoprotein and nucleic acid, and can inhibit the nucleic acid enzymolysis ability of the nucleoprotein.

A lipid nanoparticle composition for delivery of genome editing ribonucleoprotein (RNP) includes a lipid component that encapsulates the genome editing ribronucleoprotein (RNP), wherein the lipid component includes an ionizable cationic lipid with a pK a > 6, at least one phospholipid, a structural lipid, and PEG-modified lipid.

Provided is a CRISPR RNA (crRNA) applicable in a CRISPR-based ribonucleoprotein (RNP) system for reacting towards one or more genetic materials derived from Burkholderia pseudomallei in a biological sample. The crRNA comprises a spacer region having one of a poly nucleotide sequence selected from SEQ ID No. 3. SEQ ID No. 5, and SEQ ID No. 7: a repeat region preceding the spacer region forming a secondary structure thereof; and a first extension region arranged immediately after the spacer region.

Disclosed are Type V CRISPR nucleases. Related Type V CRISPR nuclease ribonucleoprotein complexes, guide nucleic acids, recombinant expression vectors, kits, methods, and systems are also provided.