30 results about "Ribonucleoprotein" 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 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.

Provided herein are self-assembled nanoparticles (NPs), pharmaceutical compositions containing such NPs and methods of using such NPs. The NPs comprise an amphiphilic copolymer and a ribonucleoprotein (RNP), and optionally ssODN, wherein: the amphiphilic copolymer is a water-soluble block copolymer comprising a poly(C2-3 alkylene glycol) block and an acrylic block comprising a poly(acrylate), poly(methacrylate) or poly(acrylate / methacrylate) block; the acrylic block comprise ester side chains bearing substituted or unsubstituted alkylamine groups; and the RNP, ssODN, and the acrylic block of the amphiphilic copolymer form a core of the self-assembled nanoparticle, and the poly(ethylene glycol) block of the amphiphilic copolymer forms the exterior of the self-assembled nanoparticle.

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 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 invention relates to improvements in reduction of tuber browning in potato (Solanum tuberosum) varieties utilising endonuclease-directed gene editing technologies In particular, the invention provides a Solanum tuberosum plant cell comprising a mutation which generates a phenotype of reduced or eliminated polyphenol oxidase activity when compared to a Solanum tuberosum plant cell without the mutation. The invention further provides ribonucleoprotein complexes comprising an endonuclease and a guide RNA that hybridises with a target sequence comprised within a polyphenol oxidase 2 (StPPO2) gene of a Solanum tuberosum variety, and methods of using the same.

The invention discloses a method for delivering a gene editing product into a fertilized egg of an animal, and belongs to the technical field of biology. The method comprises the following steps: delivering a gene editing substance into a fertilized egg of an animal by using an electroporation method, wherein the gene editing substance is a ribonucleic acid protein complex formed by a leading editing protein and RNA; and the RNA comprises pegRNA, gRNA or / or sgRNA (small guide RNA). According to the system, a RNP (ribonucleic acid protein) compound is formed by purified pilot editing protein and pegRNA transcribed in vitro, and is directly delivered to a sheep embryo by using an electroporation method. Compared with a traditional microinjection technology, the method is simple, convenient, safe and efficient. By optimizing the concentration ratio of pegRNA to PE protein in RNP, efficient editing of sheep embryo genes is realized, and the highest editing rate reaches 29.94%.

Provided herein are nuclear expressed antisense RNAs for interfering with a target repetitive RNA, the nuclear expressed antisense RNA comprising: (a) an oligonucleotide encoding an RNA-targeting guide comprising a sequence complementary to the target repetitive RNA; and (b) an oligonucleotide encoding a RNA binding protein (RBP) / ribonucleoprotein (RNP)-recruiting motif.

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 present invention relates to a method for editing the genome of a cell, such as a yeast cell. The method of the present invention requires the cell to be contacted with at least one ribonucleoprotein, at least one donor-DNA construct and a selectable marker such that they are introduced into the cell. The present invention is especially suitable for multiplex genome editing of cells such as yeast cells. The current invention further relates to a composition, a cell obtainable by the method of the invention and a method for the production of a compound of interest.

The present invention provides sesquiterpenoid derivatives and their application in the preparation of broad-spectrum antiviral drugs. [Solution] The present invention discloses sesquiterpenoid derivatives and their application in the preparation of broad-spectrum antiviral drugs, which stimulate heterogeneous nuclear ribonucleoprotein A2 / B1, activate the TANK-binding kinase 1-interferon regulatory factor 3 cell signal pathway, increase the secretion of endogenous type I interferon, and have inhibitory effects on various viruses. As a broad-spectrum antiviral drug, it can prevent or treat infectious diseases and illnesses caused by various viruses, including new coronavirus, vesicular stomatitis virus VSV-G, human immunodeficiency virus, hepatitis C virus, Japanese encephalitis virus, influenza virus, poliovirus, coxsackievirus, dengue virus, rotavirus, tobacco mosaic virus, measles virus, mumps virus, Ebola virus, Marburg virus, herpes virus and adenovirus. The sesquiterpenoid derivatives can be used as raw materials to prepare oral dosage forms such as tablets, capsules, drip pills, or clinically acceptable pharmaceutical preparations such as inhalants and injections.

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.

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.

This invention pertains to mutant Cas9 nucleic acids and proteins for use in CRISPR / Cas endonuclease systems, and their methods of use. In particular, the invention pertains to an isolated mutant Cas9 protein, wherein the isolated mutant Cas9 protein is active in a CRISPR / Cas endonuclease system, wherein the CRISPR / Cas endonuclease system displays reduced off-target editing activity and maintained on-target editing activity relative to a wild-type CRISPR / Cas endonuclease system. The invention also includes isolated nucleic acids encoding mutant Cas9 proteins, ribonucleoprotein complexes and CRSPR / Cas endonuclease systems having mutant Cas9 proteins that display reduced off-target editing activity and maintained on-target editing activity relative to a wild-type CRISPR / Cas endonuclease system.

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 polynucleotide 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.

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.

This invention pertains to mutant Lachnospiraceae bacterium ND2006 (Lb) Cas12a nucleic acids and proteins for use in CRISPR / Cas12a endonuclease systems, and their methods of use. In particular, the invention pertains to an isolated mutant LbCas12a protein, wherein the isolated mutant LbCas12a protein is active in a CRISPR / Cas12a endonuclease system. The invention also includes isolated nucleic acids encoding mutant LbCas12a proteins, ribonucleoprotein complexes and CRISPR / Cas12a endonuclease systems having mutant LbCas12a proteins.

The present article is a tabletop device for introducing a cargo(for example, a substance such as DNA, RNA, CRISPR RNPs, polymer, small molecule, carbon nanotube, protein, peptide, nanoparticle, antibody, virus, and labeling molecule)into a cell (for example, a primary blood cell, a primary immune cell, various stem cells, and a cell of an established cell line)using mechanoporation (mechanical disruption). Upon use, firstly, a cartridge holding a cell solution in which cells and the cargo are admixed is placed on the upper part of the present article. Next, the present article is pressurized and a button on the present article is pressed. Thereby, the cell solution passes through a filter within the cartridge and is pushed out into the tube placed beneath the cartridge.When the cell solution passes through the micropores of the filter, the cell membrane is mechanically disrupted, the substance permeability of the cell membrane temporarily increases, and the cargo in the cell solution is introduced into the cells. A cargo can be introduced into cells with low invasiveness and high efficiency without requiring any particular reagent or electric stimulation, and the introduced cells show high survival rates.

This invention pertains to mutant Cas9 nucleic acids and proteins for use in CRISPR / Cas endonuclease systems, and their methods of use. In particular, the invention pertains to an isolated mutant Cas9 protein, wherein the isolated mutant Cas9 protein is active in a CRISPR / Cas endonuclease system, wherein the CRISPR / Cas endonuclease system displays reduced off-target editing activity and maintained on-target editing activity relative to a wild-type CRISPR / Cas endonuclease system. The invention also includes isolated nucleic acids encoding mutant Cas9 proteins, ribonucleoprotein complexes and CRSPR / Cas endonuclease systems having mutant Cas9 proteins that display reduced off-target editing activity and maintained on-target editing activity relative to a wild-type CRISPR / Cas endonuclease system.

This invention pertains to mutant Cas9 nucleic acids and proteins for use in CRISPR / Cas endonuclease systems, and their methods of use. In particular, the invention pertains to an isolated mutant Cas9 protein, wherein the isolated mutant Cas9 protein is active in a CRISPR / Cas endonuclease system, wherein the CRISPR / Cas endonuclease system displays increased on-target editing activity relative to a wild-type CRISPR / Cas endonuclease system. The invention also includes isolated nucleic acids encoding mutant Cas9 proteins, ribonucleoprotein complexes and CRISPR / Cas endonuclease systems having mutant Cas9 proteins that display increased on-target editing activity relative to a wild-type CRISPR / Cas endonuclease system.

Presented is a nucleic acid-guided nuclease cascade assay that can detect one or more target nucleic acids of interest of interest without the need for amplifying the target nucleic acids of interest. The nucleic acid-guided nuclease cascade assays utilize signal amplification mechanisms comprising various components including nucleic acid-guided nucleases and guide RNAs (gRNAs) forming ribonucleoprotein complexes, blocked primer molecules, template molecules, polymerases and reporter moieties.

This disclosure provides compositions and methods for improving site-directed integration of nucleic acids using RNA-guided nucleases coupled with HUH endonucleases. In one aspect, this disclosure provides a ribonucleoprotein comprising: (a) a re-combinant polypeptid comprising: (i) an amino acid sequence encoding a Cas12a nuclease; (ii) an amino acid sequence encoding a linker; and (iii) an amino acid sequence encoding a HUH nuclease; and (b) at least one guide nucleic acid.

The present invention is referred to a gRNA molecule capable of recognizing the target genomic sequence located between exon 37a and exon 38 in the CACNA 1B gene, ribonucleoprotein complexes comprising said gRNA molecule, nucleic acids encoding said gRNA molecule, as well as compositions comprising them for use as a medicament, in particular in the treatment of ataxias and epilepsies.

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 belongs to the technical field of biological detection, and particularly relates to an exosome protein marker combination for cerebral small vascular diseases and related cognitive disorders and application. The invention discloses an exosome protein marker combination. The exosome protein marker combination is a combination of any one or any two or three markers of a v-type proton atp enzyme subunit, a large ribosome subunit protein eL38 and a heterogeneous ribonucleoprotein U. The exosome protein marker combination is a combination of any two or three markers of the v-type proton atp enzyme subunit, the large ribosome subunit protein eL38 and the heterogeneous ribonucleoprotein U. The combination of the three specific exosome proteins can effectively recognize CSVD related cognitive impairment, and has the potential of early warning CSVD related cognitive impairment.

The present disclosure relates to a chimeric protein molecule comprising a degradation domain including an E3 ubiquitin ligase motif without lysine residues and a targeting domain comprising a substrate-binding motif which is heterologous to the E3 ubiquitin ligase motif. A linker couples the degradation domain to the targeting domain. Also disclosed are compositions as well as methods of treating a disease, substrate silencing, forming a ribonucleoprotein, screening agents for therapeutic efficacy against a disease, and methods of screening for disease biomarkers, as well as mRNA molecules, vectors, and encapsulated nucleic acid molecules encoding chimeric protein molecules.

Compositions and methods are provided for a simple, instrumentless and sensitive method that enables rapid, instant detection of nucleic acid molecules of interest. This is based on a surprising discovery that the relative efficiency of amplification and CRISPR-based cleavage and detection can be adjusted to facilitate amplification until sufficient amplification products are generated to achieve detection. Example methods include designing a guide RNA and a primer to target a non-optimal PAM sequence, or a sequence engineered Cas nuclease to reduce its activity in forming a ribonuclein with the guide RNA or binding or cleaving a substrate nucleic acid.

The present disclosure relates to multiplexed assay methods used to detect and identify several to many target nucleic acids of interest in a sample without amplification of the target nucleic acids of interest using blocked RNP2 activator molecules that reduce or eliminate erroneous activation of ribonucleoprotein complexes in the absence of a target nucleic acid of interest.

The present invention relates to a method for editing the genome of a cell, such as a filamentous fungus cell. The method of the present invention requires the cell to be contacted with at least one pair of ribonucleoproteins, at least one donor-DNA construct and a selectable marker such that they are introduced into the cell. The present invention is especially suitable for multiplex genome editing of cells such as filamentous fungus cells. The current invention further relates to a composition, a cell obtainable by the method of the invention and a method for the production of a compound of interest. The present invention also relates to a method for high throughput transformation of a cell such as a filamentous fungus cell.

Presented is a nucleic acid-guided nuclease cascade assay that can detect one or more target nucleic acids of interest of interest without the need for amplifying the target nucleic acids of interest. The nucleic acid-guided nuclease cascade assays utilize signal amplification mechanisms comprising various components including nucleic acid-guided nucleases and guide RNAs (gRNAs) forming ribonucleoprotein complexes, blocked primer molecules, template molecules, polymerases and reporter moieties.