65 results about "Ribonucleoprotein" patented technology

The present invention relates methods of generating infectious negative-strand virus in host cells by an entirely vector-based system without the aid of a helper virus. In particular, the present invention relates methods of generating infectious recombinant negative-strand RNA viruses intracellularly in the absence of helper virus from expression vectors comprising cDNAs encoding the viral proteins necessary to form ribonucleoprotein complexes (RNPs) and expression vectors comprising cDNA for genomic viral RNA(s) (vRNAs) or the corresponding cRNA(s). The present invention also relates to methods of generating infectious recombinant negative-strand RNA viruses which have mutations in viral genes and / or which express, package and / or present peptides or polypeptides encoded by heterologous nucleic acid sequences. The present invention further relates the use of the recombinant negative-strand RNA viruses or chimeric negative-strand RNA viruses of the invention in vaccine formulations and pharmaceutical compositions.

The present invention is directed to telomerase nucleic acids and amino acids. In particular, the present invention is directed to nucleic acid and amino acid sequences encoding various telomerase protein subunits and motifs, including the 123 kDa and 43 kDa telomerase protein subunits of Euplotes aediculatus, and related sequences from Schizosaccharomyces, Saccharomyces sequences, and human telomerase. The present invention is also directed to polypeptides comprising these telomerase protein subunits, as well as functional polypeptides and ribonucleoproteins that contain these subunits.

The present invention is directed to novel telomerase nucleic acids and amino acids. In particular, the present invention is directed to nucleic acid and amino acid sequences encoding various telomerase protein subunits and motifs, including the 123 kDa and 43 kDa telomerase protein subunits of Euplotes aediculatus, and related sequences from Schizosaccharomyces, Saccharomyces sequences, and human telomerase. The present invention is also directed to polypeptides comprising these telomerase protein subunits, as well as functional polypeptides and ribonucleoproteins that contain these subunits.

Provided herein are compositions and methods useful, inter alia, for the delivery of ribonucleoprotein complexes (e.g., Cas9 / guide RNA complexes) into cells. The compositions and methods provided herein are particularly useful for the delivery of ribonucleoprotein complexes into pluripotent cells and lymphatic cells.

This invention relates to the use of tumor-derived or associated extracellular ribonucleic acid (RNA) found circulating in the plasma or serum fraction of blood for the detection, monitoring, or evaluation of cancer or premalignant conditions. Extracellular RNA may circulate as non-bound RNA, protein-bound RNA, lipid-RNA complexes, lipoprotein (proteolipid)-RNA complexes, protein-RNA complexes including within or in association with ribonucleoprotein complexes, nucleosomes, or within apoptotic bodies. Any intracellular RNA found in plasma or serum can additionally be detected by this invention. Specifically, this invention enables the extraction of circulating RNA from plasma or serum and utilizes nucleic acid amplification assays for the identification, detection, inference, monitoring, or evaluation of any neoplasm, benign, premalignant, or malignant, in humans or other animals, which might be associated with that RNA. Further, this invention allows the qualitative or quantitative detection of tumor-derived or associated extracellular RNA circulating in the plasma or serum of humans or animals with or without any prior knowledge of the presence of cancer or premalignant tissue.

The present invention is directed to novel telomerase nucleic acids and amino acids. In particular, the present invention is directed to nucleic acid and amino acid sequences encoding various telomerase protein subunits and motifs, including the 123 kDa and 43 kDa telomerase protein subunits of Euplotes aediculatus, and related sequences from Schizosaccharomyces, Saccharomyces sequences, and human telomerase. The present invention is also directed to polypeptides comprising these telomerase protein subunits, as well as functional polypeptides and ribonucleoproteins that contain these subunits.

The present invention relates methods of generating infectious negative-strand virus in host cells by an entirely vector-based system without the aid of a helper virus. In particular, the present invention relates methods of generating infectious recombinant negative-strand RNA viruses intracellularly in the absence of helper virus from expression vectors comprising cDNAs encoding the viral proteins necessary to form ribonucleoprotein complexes (RNPs) and expression vectors comprising cDNA for genomic viral RNA(s) (vRNAs) or the corresponding cRNA(s). The present invention also relates to methods of generating infectious recombinant negative-strand RNA viruses which have mutations in viral genes and / or which express, package and / or present peptides or polypeptides encoded by heterologous nucleic acid sequences. The present invention further relates the use of the recombinant negative-strand RNA viruses or chimeric negative-strand RNA viruses of the invention in vaccine formulations and pharmaceutical compositions.

The invention provides a western blot kit for detecting the antibody of autoimmune disease and a preparation method of the western blot kit, and relates to a western blot kit for detecting related antibodies of various autoimmune diseases, aiming at overcoming the technical defect that a western blot product is unavailable for testing and screening various autoimmune diseases in the prior art. The nitrocellulose membrane or the nylon membrane contains at least two parallel detection lines coated by at least two of ten natural antigens or recombinant antigens, i.e. dsDNA (deoxyribonucleic acid), Sm / RNP (ribonucleoprotein), CCP (critical compression pressure), SSA (sulfosalicylic acid), SSB (single-strand binding protein), GAD (glutamic acid decarboxylase), ICA (islet cell antibody), IA-2A (islet cell), TG (triglyceride) and AMA-M2 (anti-mitochondrial antibody), a high-concentration quality control band, a median-concentration quality control band and a low-concentration quality control band. The deficiency of the detection sensitivity and the specificity of the single autoantibody can be overcome, the operating complexity for independently detecting the related autoantibody of various diseases one by one can be overcome, and the detection efficiency and the result judging accuracy degree can be greatly improved.

This invention pertains to recombinant AsCpf1 and LbCpf1 nucleic acids and polypeptides for use in CRISPR / Cpf1 endonuclease systems and mammalian cell lines encoding recombinant AsCpf1 or LbCpf1 polypeptides. The invention includes recombinant ribonucleoprotein complexes and CRSPR / Cpf1 endonuclease systems having a suitable AsCpf1 crRNA is selected from a length-truncated AsCpf1 crRNA, a chemically-modified AsCpf1 crRNA, or an AsCpf1 crRNA comprising both length truncations and chemical modifications. Methods of performing gene editing using these systems and reagents are also provided.

The invention relates to a high-efficiency artificial activating transcription factor dCas9-TV, and a coding gene and applications thereof. According to a construction method, the carboxyl terminal ofnuclease inactivated Cas9 protein (dCas9) is connected with a plurality of copies of VP64 and TAL transcription-activating domains so as to obtain a series of novel artificial activating transcription factors, and obtain dCas9-TV with the best transcriptional activation activity via screening. When only one guide RNA (gRNA) is adopted for targeting a specific gene promoter, dCas9-TV is capable ofrealizing high efficiency activating of transcription of endogenous genes of Arabidopis thaliana and paddy rice; when a plurality of gRNA are adopted for targeting a plurality of target genes, dCas0-TV is capable of realizing transcription activation of a plurality of genes. In addition, it is confirmed that dCas9-TV possesses the same high efficiency targeting transcription activation activity in human cells. An in vitro assembled dCas9-TV / gRNA ribonucleoprotein compound can be adopted for transcription activation of Arabidopis thaliana and paddy rice endogenous genes. The high-efficiency artificial activating transcription factor dCas9-TV can be adopted in the fields such as genome genetic screening, metabolite biosynthesis pathway reconstruction, and crop improvement.

The invention discloses lupus erythematosus detection protein chip and kit thereof. The chip comprises a substrate, protein markers distributed in an array type and control point coatings, wherein the markers and the control point coatings are seven antigen markers, positive controls and negative controls comprising dsDNA (double-stranded deoxyribonucleic acid), ssDNA (single stranded deoxyribonucleic acid), Histon, SS-A(Ro60), SS-B(La), Sm (smith) and RNP (ribonucleoprotein) / Sm which are uniformly distributed on the substrate in a dot matrix mode. The invention has comprehensive and appropriate detection indexes and consistent applied reaction conditions, can detect a plurality of indexes at a time, is convenient and quick, greatly improves the detection efficiency and reduces the detection cost. The invention can be suitable for aided diagnosis of suspected lupus erythematosus disease people.

Provided are methods and compositions for delivering a nucleic acid, protein, and / or ribonucleoprotein payload to a cell. Also provided are delivery molecules that include a peptide targeting ligand conjugated to a protein or nucleic acid payload (e.g., an siRNA molecule), or conjugated to a charged polymer polypeptide domain (e.g., poly-arginine such as 9R or a poly-histidine such as 6H, and the like). The targeting ligand provides for (i) targeted binding to a cell surface protein, and (ii) engagement of a long endosomal recycling pathway. As such, when the targeting ligand engages the intended cell surface protein, the delivery molecule enters the cell (e.g., via endocytosis) but is preferentially directed away from the lysosomal degradation pathway.

The present invention relates to a method for identifying a binding site on an RNA transcript, wherein the binding site binds to one or more binding moieties. The method includes, among other things, introducing a photoreactive nucleoside into living cells wherein the living cells incorporate the photoreactive nucleoside into RNA transcripts during transcription thereby producing modified RNA transcripts; reverse transcribing the RNA of isolated cross-linked segments thereby generating cDNA transcripts with one mutation wherein the photoreactive nucleoside is transcribed to a mismatched deoxynucleoside; amplifying the cDNA transcripts thereby generating amplicons; and analyzing the sequences of the amplicons aligned against the reference sequence so as to identify the binding site, wherein the sequences of each amplicon having a mutation resulting from the introduction of the photoreactive nucleoside is considered to be a valid amplicon comprising at least a portion of a binding site on the RNA transcript.

The present invention provides a self assembly molecule having an affinity for one or more target molecules, for use in formation of a heptameric complex, comprising: a) a monomer comprising a multimerization domain of Archaeal Sm1 (AF-Sm1) protein or SM-like ribonucleoprotein from other organisms, able to interact with other molecules of the same monomer comprising a multimerization domain of AF-Sm1 protein or SM-like ribonucleoprotein to self-assemble into a heptamer; and b) a target binding domain or peptide attached directly or via a linker to the monomer of (a). Also provided are heptamers comprising these self assembly molecules and methods for their use in therapy, imaging and diagnostics.

This invention relates to the use of tumor-derived or associated extracellular ribonucleic acid (RNA) found circulating in the plasma or serum fraction of blood for the detection, monitoring, or evaluation of cancer or premalignant conditions. Extracellular RNA may circulate as non-bound RNA, protein-bound RNA, lipid-RNA complexes, lipoprotein (proteolipid)-RNA complexes, protein-RNA complexes including within or in association with ribonucleoprotein complexes, nucleosomes, or within apoptotic bodies. Any intracellular RNA found in plasma or serum can additionally be detected by this invention. Specifically, this invention enables the extraction of circulating RNA from plasma or serum and utilizes nucleic acid amplification assays for the identification, detection, inference, monitoring, or evaluation of any neoplasm, benign, premalignant, or malignant, in humans or other animals, which might be associated with that RNA. Further, this invention allows the qualitative or quantitative detection of tumor-derived or associated extracellular RNA circulating in the plasma or serum of humans or animals with or without any prior knowledge of the presence of cancer or premalignant tissue.

This invention relates to the use of tumor-derived or associated extracellular ribonucleic acid (RNA) found circulating in the plasma or serum fraction of blood for the detection, monitoring, or evaluation of cancer or premalignant conditions. Extracellular RNA may circulate as non-bound RNA, protein-bound RNA, lipid-RNA complexes, lipoprotein (proteolipid)—RNA complexes, protein-RNA complexes including within or in association with ribonucleoprotein complexes, nucleosomes, or within apoptotic bodies. Any intracellular RNA found in plasma or serum can additionally be detected by this invention. Specifically, this invention enables the extraction of circulating RNA from plasma or serum and utilizes nucleic acid amplification assays for the identification, detection, inference, monitoring, or evaluation of any neoplasm, benign, premalignant, or malignant, in humans or other animals, which might be associated with that RNA. Further, this invention allows the qualitative or quantitative detection of tumor-derived or associated extracellular RNA circulating in the plasma or serum of humans or animals with or without any prior knowledge of the presence of cancer or premalignant tissue.

The present application provides the amino acid and nucleic acid sequences of heavy and light chain complementarity determining regions of a cancer specific antibody directed to an epitope of variant Heterogeneous Ribonucleoprotein G (HnRNPG). In addition, the application provides cancer specific antibodies and immunoconjugates comprising the cancer specific antibody attached to a toxin or label, and methods of uses thereof. The application also relates to diagnostic methods and kits using the cancer specific antibodies disclosed herein. Further, the application provides novel cancer-associated epitopes and antigens of variant HnRNPG, and uses thereof.

The invention discloses a barley gingko milk product and a preparation method thereof. Barley sprout powder, gingko, pure milk or milk powder are used as main raw materials, through the scientific formula and the advanced process, the barley gingko milk product is produced, and has faint scent taste, rich nutrition and higher health function. The product not only has components of crude protein, crude fat, reducing sugar, ribonucleoprotein, mineral substance, crude fiber, multiple vitamins and the like, but also has better health functions of improving acidic constitution and preventing various diseases.

The invention relates to preparation and application of an enzyme-linked immuno sorbent assay (ELISA) kit for detecting a virus-specific antibody of fever with throbocytopenia associated syndrome. The kit comprises an anti-human-IgM-enveloped ELISA reaction plate, horse radish peroxidase (HRP)-coupled nucleocapsid protein (NP) and a tetramethylbenzidine (TMB) color development agent. A preparation method for a ribonucleoprotein (rNP)-enveloped reaction plate comprises the following steps of: amplifying an open reading frame sequence of nucleocapsid protein of a virus strain of a novel bunyavirus JS-2007-001 by a reverse transcription polymerase chain reaction (PCR) method, inserting the sequence into pQE30 to construct a pQE30-NP expression plasmid, transfecting an M15 strain by using pQE30-NP, performing inducible expression of 6His-NP recombinant protein by using isopropyl-beta-d-thiogalactoside (IPTG), purifying the 6His-NP recombinant protein by using Hitrap, and marking to obtain HRP-coupled NP(HRP-rNP). The reaction plate can be combined with a novel bunyavirus-resistant antibody (IgM) in a sample when incubated with the sample to be tested conventionally, and the HRP-rNP incubation is used in the subsequent detection, so that an 'anti-human IgM-antibody-enzyme-labeled antigen' capture detection system is formed; the color reaction after an enzyme substrate, namely the TMB color development agent is added is analyzed by an enzyme-labeled instrument; and the content of the specific antibody (IgM) of the novel bunyavirus is obtained according to the color development degree.

Provided herein are nanocapsules comprising a single ribonucleoprotein (RNP) complex as a core and an biodegradable crosslinked polymer shell that encapsulates the core, wherein the RNP complex comprises a Cas9 polypeptide and a guide RNA, and the biodegradable crosslinked polymer shell comprises polymerized monomers of imidazolyl acryloyl monomers, bisacryloyl disulfide monomers (a biodegradable cross-linker), optionally PEG acryloyl monomers, and either cationic acryloyl monomers, anionic acryloyl monomers, or both cationic and anionic acryloyl monomers (optionally in combination with non-ionic acryloyl monomers) as defined herein. Also provided are methods of making the nanocapsules, kits containing the nanocapsules and methods of delivering the encapsulated RNP to cells.

The invention discloses an efficient clustered regularly interspaced short palindromic repeats ribonucleoprotein (CRISPR RNP) and donor DNA co-location mediated gene insertion or replacement method and application thereof. The method comprises the following steps: (1) biotin labeling is performed on donor DNA to obtain biotin-labeled donor DNA; (2) fusion protein of Cas9 protein and monovalent streptavidin, sgRNA and the biotin-labeled donor DNA are evenly mixed and undergo standing to obtain a CRISPR RNP-donor DNA complex; and (3) cell nucleus transformation is performed on the CRISPR RNP-donor DNA complex to realize gene insertion or replacement. According to the method, by utilizing the characteristic that the fusion protein can be combined with the biotin-labeled donor DNA, the CRISPRRNP and the donor DNA appear at a target locus together, and precise insertion of the donor DNA at the target locus or accurate replacement of a gene at the target locus is realized; and the method issuitable for the aspect of crop and livestock breeding.

The invention discloses a kit (chemiluminiscence method) for determining an anti-ribonucleoprotein 70 antibody IgG. The kit comprises magnetic particles coated by anti-ribonucleoprotein 70 antigen, acridinium ester marked by mouse-anti-human IgG, a sample diluent, a test diluent, an anti-ribonucleoprotein 70 antibody IgG calibration product, pre-exciting liquid and exciting liquid. The invention further discloses a manufacturing method of the kit (chemiluminiscence method) for determining the anti-ribonucleoprotein 70 antibody IgG. Compared with an existing kit, the kit has the advantages of being easy to operate, high in sensitivity and wide in detection range.

The present invention describes vesicles carrying gRNA(s) and / or CRISPR-associated RNA guided-nuclease ribonucleoprotein complexes (RNPs) that are efficiently delivered into target cells through fusogenic envelope proteins.