46 results about "Argonaute" patented technology

A method for producing a plant with increased yield as compared to a corresponding wild type plant whereby the method comprises at least the following step: increasing or generating in a plant or a part thereof one or more activities of a polypeptide selected from the group consisting of 2-oxoglutarate-dependent dioxygenase, 3-ketoacyl-CoA thiolase, 3′-phosphoadenosine 5′-phosphate phosphatase, 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase, 5OS chloroplast ribosomal protein L21, 57972199. R01.1-protein, 60952769. R01.1-protein, 60S ribosomal protein, ABC transporter family protein, AP2 domain-containing transcription factor, argonaute protein, AT1 G29250.1-protein, AT1 G53885-protein, AT2G35300-protein, AT3G04620-protein, AT4G01870-protein, AT5G42380-protein, AT5G47440-protein, CDS5394-protein, CDS5401_TRUNCATED-protein, cold response protein, cullin, Cytochrome P450, delta-8 sphingolipid desaturase, galactinol synthase, glutathione-S-transferase, GTPase, haspin-related protein, heat shock protein, heat shock transcription factor, histone H2B, jasmonate-zim-domain protein, mitochondrial asparaginyl-tRNA synthetase, Oligosaccharyltransferase, OS02G44730-protein, Oxygen-evolving enhancer protein, peptidyl-prolyl cis-trans isomerase, peptidyl-prolyl cis-trans isomerase family protein, plastid lipid-associated protein, Polypyrimidine tract binding protein, PRLI-interacting factor, protein kinase, protein kinase family protein, rubisco subunit binding-protein beta subunit, serine acetyltransferase, serine hydroxymethyltransferase, small heat shock protein, S-ribosylhomocysteinase, sugar transporter, Thioredoxin H-type, ubiquitin-conjugating enzyme, ubiquitin-protein ligase, universal stress protein family protein, and Vacuolar protein.

The invention provides a nucleic acid testing method based on a prokaryotic Argonaute protein and an application of the nucleic acid testing method, and particularly provides a system for detecting atarget nucleic acid molecule. The system comprises guide ssDNAs, gene editing enzyme Pyrococcus furiosus Argonaute (PfAgo) and a fluorescence reporter nucleic acid. The nucleic acid testing method ishigh in sensitivity, good in specificity and high in flux and can be widely applied to the fields of molecular medical diagnosis, food safety testing, environmental monitoring and the like.

Methods of developing genetically engineered immune cells for immunotherapy, which can be endowed with Chimeric Antigen Receptors targeting an antigen marker that is common to both the pathological cells and said immune cells (ex: CD38, CS1 or CD70) by the fact that the genes encoding said markers are inactivated in said immune cells by a rare cutting endonuclease such as TALEN, Cas9 or argonaute.

Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome ofa cell or subject, e. g., within the human genome. In some embodiments, fusion proteins of nucleic acid programmable DNA binding proteins (napDNAbp), e. g., Cpf 1 or variants thereof, and nucleic acid editing proteins or protein domains, e. g., deaminase domains, are provided. In some embodiments, methods for targeted nucleic acid editing are provided. In some embodiments, reagents and kits for the generation of targeted nucleic acid editing proteins, e. g., fusion proteins of a napDNAbp (e. g., CasX, CasY, Cpfl, C2cl, C2c2, C2C3, and Argonaute) and nucleic acid editing proteins or domains, are provided.

The present invention relates to an Argonaute protein mutant which lacks DNA cleavage activity, but has DNA binding activity. The mutation of the mutant is located in a PIWI domain. The invention alsorelates to application of the protein mutant particularly in the enrichment of target DNA and in the construction of a sequencing library. Therefore, the present invention also relates to a method for enriching the target DNA. The method comprises the steps of: (a) designing a leader sequence for a specific sequence in the target DNA; (b) combining the mutant, the leader sequence and the target DNA according to the present invention to obtain a mutant-leader sequence-target DNA ternary complex; (c) capturing the mutant-leader sequence-target DNA ternary complex through a capture medium; and (d) isolating the target DNA from the captured mutant-leader sequence-target DNA ternary complex to obtain enriched target DNA.

The present application provides methods, compositions, delivery systems, and kits for modifying a target nucleic acid using an Argonaute (Ago) and a single-stranded guide DNA. In some embodiments, methods of gene editing in a cell, such as a mammalian cell, are provided.

This invention relates to a method to produce gene alterations in the genomes of a eukaryotic and prokaryotic host cell. The method comprises utilizing Argonaute from Natronobacterium gregoryi (NgAgo) or a mutant thereof, and complementary 5′ phosphorylated single-stranded DNA that target the enzyme to cleave specific regions of the chromosome. Additionally, N-terminal truncations (deletion of the repA domain; N-del), or its mutants including N-del / E598A, N-del / D601P, and N-del / E602P reduces random cleavage and can be used for targeted gene editing with a guide DNA. An expression system or a host cell and method of creating thereof are also in the scope of this application.

The invention discloses an Argonaute protein derived from eukaryotes and application of the Argonaute protein. The amino acid sequence of the Argonaute protein is as shown in SEQ ID NO.1, or the amino acid sequence of the Argonaute protein has at least 50% of sequence consistency with the sequence as shown in SEQ ID NO.1; according to the invention, the specific cleavage activity of the eukaryotic Argonaute protein on DNA is proved for the first time, and experimental proof is provided for research on interaction between the eukaryotic Argonaute and DNA; meanwhile, the polypeptide, the nucleic acid, the expression vector, the composition, the kit and the method used in the invention can perform site-specific operation on intracellular and extracellular genetic materials, can be effectively applied to many fields of biotechnology, and provide a new tool for gene editing, modification and molecular detection based on the Argonaute polypeptide derived from eukaryotes.

The invention discloses a gene editing application of a prokaryotic Argonaute protein, which comprises the following steps: constructing a prokaryotic expression vector of the Argonaute protein derived from bacteria, detecting the degradation of the expression vector and a co-transformation vector of the Argonaute protein after transforming a host cell, and detecting genome sites. It is found that the gene has the ability of being activated by a specific transcript and targeting a specific DNA target sequence, and gene editing with the inDel frequency of 3% is carried out on a genome target sequence under the participation of guide RNA transcribed in eukaryotic cells. Experimental results show that the prokaryotic Argonaute protein can be used for developing intracellular gene editing tools.

Methods of developing genetically engineered immune cells for immunotherapy, which can be endowed with Chimeric Antigen Receptors targeting an antigen marker that is common to both the pathological cells and said CD38 immune by the fact that the genes encoding said markers are inactivated in said immune cells by a rare cutting endonuclease such as TALEN, Cas9 or argonaute.

A composition for treating a lysogenic virus, including isolated nucleic acid encoding two or more gene editors chosen from gene editors that target viral DNA, gene editors that target viral RNA, andcombinations thereof. A composition for treating a lytic virus, including isolated nucleic acid encoding at least one gene editor that targets viral DNA and a viral RNA targeting composition. A composition for treating both lysogenic and lytic viruses, including isolated nucleic acid encoding two or more gene editors that target viral RNA, chosen from CRISPR-associated nucleases, Argonaute endonuclease gDNAs, C2c2, RNase P RNA, and combinations thereof. A composition for treating lytic viruses, including isolated nucleic acid encoding two or more gene editors that target viral RNA and a viralRNA targeting composition. Methods of treating a lysogenic virus or a lytic virus, by administering the above compositions to an individual having a virus and inactivating the virus.

The present disclosure provides compositions, kits, genetically modified cells, non-human transgenic organisms, and methods for binding and / or cleaving a single stranded target nucleic acid. A method of cleaving includes contacting a single stranded target nucleic acid with (e.g., introducing into a cell) a subject argonaute (Ago) polypeptide and a guide RNA (e.g., having a 5′-OH). In some embodiments, a subject Ago polypeptide includes an amino acid sequence having 70% or more sequence identity with amino acids 282-430 and / or 431-639 of the Marinitoga piezophila argonaute (MpAgo) protein set forth in SEQ ID NO: 1. The present disclosure provides variant Ago polypeptides; and methods of use of same.

The invention provides characterization and application of a novel high-temperature Argonaute protein. Specifically, the invention provides a nucleic acid cleavage system, which is characterized in that the nucleic acid cleavage system comprises: (a) guide DNA (gDNA); (b) a programmable endonuclease Argonaute (Ago); and (c) optionally a reporter nucleic acid, wherein if the reporter nucleic acid is cleaved, the cleavage can be detected. In addition, the invention also provides a system and a method for enriching and detecting low-abundance target nucleic acid based on the programmable endonuclease TpsAgo provided by the invention. The detection system and method provided by the invention can specifically and effectively enrich and efficiently detect low-abundance nucleic acid, and the programmable endonuclease TpsAgo provided by the invention has strong gene manipulation potential.

The invention provides a prokaryotic Argonaute protein-based nucleic acid detection method and application thereof. Specifically, the present invention provides a detection system for detecting target nucleic acid molecules, the system comprising guide ssDNAs, gene editing enzyme Pyrococcus furiosus Argonaute (PfAgo) and fluorescent reporter nucleic acid. The nucleic acid detection method of the invention has high sensitivity, good specificity, and high throughput, and can be widely used in many fields such as molecular medical diagnosis, food safety detection, and environmental monitoring.

The invention relates to a method for eliminating a self-connector of a sequencing library and an application. The method comprises the following steps that a short sequence guide DNA is designed by taking a sequencing connector self-connecting library sequence as a target sequence, double-strand breakage of a dsDNA library molecule is realized by combining with Argonaute endonuclease, and the self-connector is cut from the dsDNA library molecule, so as to prevent the self-connector from being amplified in a subsequent PCR reaction. According to the method for eliminating the self-connector of the sequencing library, the proportion of self-connectors of the library can be remarkably reduced, the sequencing clean reads can be increased, and the data efficiency can be improved.