175 results about "Nucleoprotein" patented technology

The technology described herein relates to novel CRISPR / Cas systems and components thereof, vectors, proteins, fusion proteins, ribonucleoprotein complexes and methods of using the novel systems and components. The new system can be used in CRIS PRi applications and can be fused with any number of effector domain patterns, such as nuclease, base editor, leader editor, and the like, to conduct a variety of DNA modifications.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody combination for detecting measles virus nucleoprotein and application of the monoclonal antibody combination. The combination is composed of monoclonal antibodies 5E2 and 5G9 and can efficiently and specifically recognize measles virus nucleoprotein, amino acid sequences of complementary determining regions (CDR) of heavy chain and light chain variable regions of the combination are clear and are shown as SEQ ID NO.1-12 respectively, high specificity and high affinity of the antibodies are guaranteed, the antibody combination can effectively capture and detect target antigens, and the antibody combination can be used for detecting measles virus nucleoprotein. The risk of non-specific binding and cross reaction is obviously reduced. The combination shows excellent sensitivity and specificity in detection platforms such as colloidal gold immunochromatography, and provides a stable and reliable biological recognition tool for rapid and accurate detection of measles viruses.

Disclosed are methods of treating a subject with a neurological disease associated with a splicing defect caused by TDP-43 proteinopathies, comprising administering to said subject an agent to increase expression levels and / or stability of hnRNP L, thereby attenuating and / or repairing the splicing defect. The disclosure also relates to nucleic acids targeting heterogeneous nuclear ribonucleoprotein L (hnRNP L), and their use.

The invention relates to a plasma biomarker for predicting the curative effect of non-small cell lung cancer immunotherapy and application of the plasma biomarker. The biomarker for predicting the curative effect of the non-small cell lung cancer immunotherapy, disclosed by the invention, comprises phosphoglycerate kinase 1 (PGK1), fibronectin 1 (FN1), chromogranin A (CHGA), elastin (ELN), heterogeneous ribonucleoprotein H3 (HNRNPH3) and an Aly / REF output factor (ALYREF). The plasma biomarkers for predicting the non-small cell lung cancer immunotherapy curative effect can be detected by adopting a mass spectrum method, and the non-small cell lung cancer immunotherapy curative effect of a patient is judged by the markers.

The invention belongs to the technical field of biological pharmacy, and particularly relates to influenza virus-like particles for chimeric expression of new coronavirus RBD protein as well as a preparation method and application of the influenza virus-like particles. The virus-like particle is prepared by embedding fusion protein on the surface of influenza virus M1, and the fusion protein comprises an HA2 segment of HA protein of influenza virus, an extracellular functional region of M2e matrix protein, a conserved region short peptide of nucleoprotein, and RBD tandem protein with mutated five sites of 417, 452, 478, 484 and 501 of new coronavirus; the virus-like particles can be used for preventing specific influenza and new crowns; the preparation method of the virus-like particles is simple and low in cost, the virus-like particles are good in immunogenicity and cross protectiveness, the preparation requirements of large-scale influenza vaccines can be met, the biological safety is high, and the immunity of an organism to various influenza viruses and new coronal viruses can be stimulated.

The invention provides a chimeric bivalent SFTSV (severe fever with thrombocytopenia syndrome virus) mRNA (messenger ribonucleic acid) vaccine, which is characterized in that the vaccine co-expresses an SFTSV envelope protein precursor GPC and a nucleoprotein NP in series through a flexible joint. Through humoral immunity, cellular immunity and challenge protection effect evaluation, the effectiveness of the mRNA vaccine provided by the invention is verified. Research results show that the prepared SFTSV mRNA vaccine can generate a good immune protection effect by only needing one mRNA, has the characteristics of simple industrial production, easy quality control and the like, and is an ideal choice of the SFTSV vaccine.

The invention provides a humanized monoclonal antibody for detecting a human-synuclein oligomer and application of the humanized monoclonal antibody, and belongs to the technical field of biological engineering. The light chain CDR sequence of the binding part comprises at least one of LCDR1, LCDR2 and LCDR3, and the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 1; the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 2, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 3. The monoclonal antibody is used as a raw material, and a unique repeated epitope double-antibody sandwich immunodetection method is adopted, so that the human a-Syn oligomer prepared in vitro or in a sample can be detected with high specificity, high sensitivity and high accuracy.

Provided herein are SNCA antisense oligonucleotides that are capable of modulating the expression of a SNCA target nucleic acid. Also provided herein are methods of use thereof.

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 a TIMM10 gene as a target spot in resisting influenza A virus, and belongs to the technical field of biological medicine. Specifically, TIMM10 gene knock-down is carried out on A549 cells, and it is found that in the cells with knocked-down TIMM10, expression of main protein hemagglutinin and nucleoprotein of influenza viruses is remarkably improved, and virus RNA and virus titer in supernate are also remarkably increased; overexpression of TIMM10 in A549 cells inhibits replication and infectivity of IAV in the cells. A further in-vivo experiment is carried out through a TIMM10 whole body knockout heterozygote mouse, and a result shows that the lung of the TIMM10 heterozygote mouse shows more serious pulmonary infection, including pulmonary hemorrhage and increased inflammatory cell infiltration. Based on the effect of TIMM10 in the influenza virus infection process, the invention provides a new therapeutic target, and provides a new thought and direction for developing a therapeutic strategy for IAV in the future.

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.

Systems and methods for highly-effective CRISPR-Cas based genomic editing within human chromosome 15q11-q13 have been developed as therapeutic interventions for Angelman Syndrome (AS). Selective single guide RNA molecules (sgRNAs) that impart enhanced CRISPR-Cas editing of genomic mutations associated with Angelman Syndrome in human cells are described. The engineered sgRNAs induce activity of non-pathogenic, paternal UBE3A alleles to reduce, reverse and / or prevent the causative neurodevelopmental defects of AS in a subject in need thereof. Compositions and methods of engineered crRNAs, sgRNAs thereof and ribonucleoprotein (RNP) complexes thereof are provided for enhanced genomic engineering with increased on-off target specificity and on-target editing efficacy for treatment of AS.

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 invention discloses an efficient extraction method of blue fox sperm DNA, and belongs to the technical field of molecular biology. Aiming at the structural characteristics that chromatin of the blue fox sperms is highly condensed, histone is replaced by protamine, and a stable nucleoprotein complex is formed through an intermolecular disulfide bond, the invention provides an efficient DNA extraction method suitable for the blue fox sperms. According to the method, based on the synergistic effect of three chemical reagents, namely SDS, PK and DTT, full lysis of the sperm cells of the blue foxes is achieved, and then efficient enrichment of sperm DNA of the blue foxes can be achieved by combining a conventional DNA extraction method. According to the efficient extraction method of the blue fox sperm DNA provided by the invention, the yield and integrity of the blue fox sperm DNA are remarkably improved, and the limitation that sperm chromatin is difficult to effectively lyse by a conventional method is overcome; the method provides reliable technical support for research on genetic diversity evaluation, population management, molecular breeding and the like of the blue foxes, and has the potential of popularization and application in other high-condensation sperm species.

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 invention discloses an ultra-sensitive target nucleic acid enrichment detection method based on programmable nuclease, and belongs to the technical field of biology. According to the enrichment method disclosed by the invention, on the basis of specific cutting of programmable nuclease, wild type nucleic acid can be continuously removed while isothermal amplification is carried out, so that the number of mutant alleles in a sample is continuously and exponentially increased, and low-abundance mutant genes or methylated DNA are enriched or detected. According to the present invention, sgRNA, guide DNA or guide RNA are designed and optimized, the sgRNA, the guide DNA or the guide RNA and a Cas protein or an Ago protein are used to form a ribonucleoprotein complex, the isothermal amplification is performed while the specific cutting is performed on the target gene, the mutation-containing gene is not cut, and the low-abundance mutation gene is enriched;

The invention relates to the field of aquaculture, in particular to a preparation method and application of a recombinant live vector vaccine for infectious hematopoietic necrosis of fish. In order to solve the problem of prevention and control of the infectious hematopoietic necrosis of the fish in the aquatic product industry, a replication-defective adenovirus type 5 is selected as a target gene expression vector to construct a recombinant adenovirus vector comprising a glycoprotein gene and a nucleoprotein gene of the infectious hematopoietic necrosis virus of the fish, and a recombinant virus is obtained; the safe and effective recombinant live vector vaccine for the infectious hematopoietic necrosis of the fish is created. According to the vaccine, immunization is carried out on fishes in a soaking or bait feeding mode, the total CPM of the fishes immunized by the vaccine is 6.67%, and the total CPM of the fishes in a non-immunized control group is 94.00%. In addition, the RPS of the vaccination group fish and the RPS of the empty vector control group fish are 92.90% and 9.22% respectively. The method is at a leading level in the field of fish biological medicines at present and has remarkable creativity.

The present application relates to a polynucleotide construct for gene editing suitable to hybridize with a CRISPR-Cas9 ribonucleoprotein complex. The disclosed polynucleotide construct is suitable for the spatially controlled release of CRISPR-Cas9 ribonucleoprotein for gene editing. The gene editing potential of the construct in a composition was demonstrated in vitro at single cell level. The safety and gene editing of the construct was also demonstrated. This construct offers a new strategy for the delivery of the CRISPR system with spatial control.

The present application relates to the technical field of biotechnology, and particularly relates to a nanocomposite containing a cationic lipid analogue and application of the cationic lipid analogue in intracellular delivery of a gene editing ribonucleoprotein complex. The nanocomposite comprises the cationic lipid analogue and the ribonucleoprotein complex; the cationic lipid analogue has a structure as shown in formula (I). The nanocomposite has a high gene editing effect on different gene sites (AAVS1, HBB, EGFP and KRAS) in tumor tissues, which further indicates that the cationic lipid analogue delivering the ribonucleoprotein complex has a certain inhibitory effect on tumor growth; the gene editing ribonucleoprotein complex delivery carrier can achieve a high delivery efficiency in intracellular delivery and is safe and effective.

The invention relates to novel oligonucleotides and their use in inhibiting or reducing senescence. The oligonucleotides target specific sequences in the 3' untranslated region (UTR) of heterogeneous nuclear ribonucleoprotein D (HNRNPD) mRNA and blocks one or more inhibitory microRNA (miRNA) binding sites. This promotes the physiologically regulated expression of HNRNPD and reduces senescence.

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.

The invention discloses an ultra-sensitive target nucleic acid enrichment detection method based on programmable nuclease, and belongs to the technical field of biology. According to the enrichment method disclosed by the invention, on the basis of specific cutting of programmable nuclease, wild type nucleic acid can be continuously removed while isothermal amplification is carried out, so that the number of mutant alleles in a sample is continuously and exponentially increased, and low-abundance mutant genes or methylated DNA are enriched or detected. According to the present invention, sgRNA, guide DNA or guide RNA are designed and optimized, the sgRNA, the guide DNA or the guide RNA and a Cas protein or an Ago protein are used to form a ribonucleoprotein complex, the isothermal amplification is performed while the specific cutting is performed on the target gene, the mutation-containing gene is not cut, and the low-abundance mutation gene is enriched;

The present invention relates to a novel analytical method for quantifying the concentration of uncomplexed ("free") gRNA present in a Cas-gRNA ribonucleoprotein complex produced for genome editing.

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.

The invention belongs to the technical field of plant breeding, and discloses a breeding method for improving the mutagenesis efficiency of stem tips of woody plants. The CRISPR-Cas9 ribonucleoprotein complex and plant hormones 6-benzyladenine, gibberellin and jasmonic acid are co-encapsulated and then accurately delivered to stem tip meristem of woody ornamental plants such as acer truncatum, pteroceltis tatarinowii, crape myrtle and tassel, and the gene mutation obtaining rate of the stem tip meristem is effectively increased; the intelligent environment control system is used for dynamically optimizing illumination, temperature, humidity and oxidative stress, activating a plant endogenous DNA repair pathway and further enhancing homologous recombination efficiency, meanwhile, through collaborative optimization of induction hormone signals and environmental stress, the stress resistance is improved, the leaf color, the plant type and the growth vigor can be accurately regulated and controlled, and the yield of the plant is increased. According to the method, efficient, accurate and stress-resistant linked woody plant stem tip mutation breeding is realized.

The present application relates to a polynucleotide construct for gene editing suitable to hybridize with a CRISPR-Cas9 ribonucleoprotein complex. The disclosed polynucleotide construct is suitable for the spatially controlled release of CRISPR-Cas9 ribonucleoprotein for gene editing. The gene editing potential of the construct in a composition was demonstrated in vitro at single cell level. The safety and gene editing of the construct was also demonstrated. This construct offers a new strategy for the delivery of the CRISPR system with spatial control.

The invention provides a multivalent Ebola virus broad-spectrum mRNA vaccine as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. According to the invention, a vaccine is prepared on the basis of four mRNAs for expressing Ebola virus antigen proteins and lipid nanoparticles, and the antigen proteins are Zaire type Ebola virus glycoprotein, Bandong type Ebola virus glycoprotein, Sudan type Ebola virus glycoprotein and Zaire type Ebola virus nucleoprotein; preferably, the amino acid sequences of the antigen protein are shown as SEQ ID NO: 8, 9, 10 and 11, and the nucleotide sequences of the DNA of the antigen protein are shown as SEQ ID NO: 3, 5, 6 and 7. The vaccine induces strong cellular immune and humoral immune response, obtains comprehensive and crossed immune protection, and provides wide protection for various Ebola viruses, so that the vaccine can be used as a medicine for preventing Ebola virus outbreak.