2527 results about "CRISPR" patented technology

A federated distributed computational system enables secure biological data analysis and genomic medicine with enhanced oncological therapy capabilities. The system implements patient-specific tumor-on-a-chip analysis through microfluidic control systems and cellular heterogeneity preservation, while integrating fluorescence-enhanced diagnostics using CRISPR-LNP targeting and robotic surgical navigation. The architecture coordinates spatiotemporal analysis of gene therapy delivery through molecular imaging and immune response tracking, and implements bridge RNA integration with multi-target synchronization. Treatment selection is optimized through multi-criteria scoring and patient-specific simulation modeling. Each federated node contains a local processing unit for biological data analysis, privacy preservation protocols, and a hierarchical knowledge graph structure. The system implements cross-species genetic analysis, environmental response modeling, and multi-scale tensor-based data integration, enabling research institutions to collaborate on complex, large-scale biological analyses while maintaining strict data privacy controls.

The invention relates to the improvement of endonuclease-based antimicrobials by blocking DNA repair of double-strand break(s) (DSB(s)) in prokaryotic cells. In this respect, the invention especially concerns a method involving blocking DNA repair after a nucleic acid has been submitted to DSB, in particular by a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) associated programmable double-strand endonuclease. The invention particularly relates to the use of an exogenous molecule that inhibits DNA repair, preferably a protein that binds to the ends of the double-stranded break to block DSB repair. The invention also relates to vectors, particularly phagemids and plasmids, comprising nucleic acids encoding nucleases and Gam proteins, and a pharmaceutical composition and a product containing these vectors and their application.

Provided herein are gene editing pairs comprising an engineered protein, e.g, an engineered CRISPR protein and a guide ribonucleic acid (gRNA) that may provide improved gene editing relative to reference, i.e., wild type sequences. In some embodiments, the engineered protein of the gene editing pair comprises a RuvC cleavage domain with one or more amino acid modifications relative to a reference CRISPR protein RuvC sequence. In some embodiments, a guide ribonucleic acid (gRNA) of the gene editing pair comprises at least one modification in a scaffold stem loop relative to a reference scaffold stem loop sequence.

The invention discloses construction and optimization of a gene editing tool based on CRISPR (clustered regularly interspaced short palindromic repeats)-associated transsposases and application of the gene editing tool in escherichia coli, and belongs to the field of synthetic biology and gene engineering. According to the invention, an efficient gene editing system is established in Escherichia coli, and the burden of a MUSCULAR-CAST system on strains is reduced through element optimization; by eliminating the self-targeting effect, large-fragment (> 10k bp) target gene expression cassettes can be efficiently integrated, and the editing degree of the gene editing system is improved. According to the invention, a MUSCULAR-CAST gene editing system is used for constructing the chassis strains for producing 3-fucosyllactose, L-tryptophan, 2 '-fucosyllactose and neotetraose, so that the chassis strains have equivalent or higher expression yield compared with plasmids, and the growth condition is better or the conversion rate is higher.

The invention belongs to the technical field of pathogenic microorganism detection, and particularly relates to a sequence combination for detecting cochliobolus miyabeanus based on RPA-CRISPR / Cas12b and application of the sequence combination. The CRISPR / Cas system contained in the composition comprises Cas12b and sgRNA, and the CRISPR / Cas system can be cooperatively used with a specific reaction system. The rapid detection method comprises the following steps: sample pretreatment: treating a sample to extract nucleic acid; performing amplification reaction: performing RPA amplification reaction on the extracted nucleic acid by using the composition, and applying a nucleic acid amplification product to CRISPR / Cas system detection; result interpretation: reading and judging a detection result through a quality control line and a detection line. The composition and the kit provided by the invention are specially used for detecting the cochliobolus miyabeanus. The method is simple and convenient to operate, rapid in detection, short in time consumption, low in cost and suitable for on-site rapid detection.

The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing and analyte characterization. Such polynucleotide processing may be useful for a variety of applications, including analyte characterization by polynucleotide sequencing. The compositions, methods, systems, and devices disclosed herein generally describe barcoded oligonucleotides, which can be bound to a bead, such as a gel bead, useful for characterizing one or more analytes including, for example, protein (e.g., cell surface or intracellular proteins), genomic DNA, and RNA (e.g., mRNA or CRISPR guide RNAs). Also described herein, are barcoded labelling agents and oligonucleotide molecules useful for “tagging” analytes for characterization.

The invention belongs to the technical field of in-vitro detection, and particularly relates to a nucleic acid combination, a kit and a method for detecting monkey pox virus based on a CRISPR (clustered regularly interspaced short palindromic repeats) technology. The detection reagent provided by the invention comprises an RPA freeze-drying reagent and a CRISPR / Cas12a detection system freeze-drying reagent, the RPA freeze-drying reagent comprises an RPA isothermal amplification reagent, Buffer A, a front primer, a rear primer and a mixed freeze-drying protective agent; the CRISPR / Cas12a detection system freeze-drying reagent comprises Cas12a, gRNA (guide ribonucleic acid), ssDNA (single-stranded deoxyribonucleic acid) reporter molecules, Buffer 2.1 and a mixed freeze-drying protective agent. The composition of the mixed freeze-drying protective agent is optimized according to the composition of a detection system and a specific detection method. The kit solves the problems of normal-temperature storage and transportation of RPA / CRISPR detection reagents of the monkey pox virus, has the characteristics of simplicity and convenience in operation, rapidness in detection, high sensitivity, suitability for on-site and large-scale screening and the like, provides an efficient and accurate solution for large-scale screening, and has a very good application prospect.

The invention discloses a visual detection method of New Delhi tomato leaf curl virus, and belongs to a plant virus detection technology in the field of biotechnology. The method comprises the following steps: adding genome DNA extracted from a plant sample to be detected into an RAA reaction system containing an RAA primer, and carrying out isothermal amplification; then adding an isothermal amplification product into a CRISPR / Cas12a detection system containing a specific crRNA fluorescent probe and an ssDNA fluorescent probe, and carrying out a reaction; and finally, carrying out fluorescence signal detection on the reaction product, if the fluorescence signal is positive, indicating that the New Delhi tomato leaf curl virus is detected in the plant sample, otherwise, indicating that the New Delhi tomato leaf curl virus is not detected. The method has the advantages of being good in specificity, short in detection time, high in sensitivity, free of dependence on expensive instruments and the like, and the result can be accurately detected through an instrument or directly and preliminarily interpreted in a field with limited conditions.

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The invention discloses an integrated fluorescence detection kit and a testing method thereof.The integrated fluorescence detection kit comprises an upper cover assembly, a lysate tube assembly, a main body assembly, a diluent tube assembly and a decorative cover, and the lower end of the upper cover assembly is connected with the lysate tube assembly and the diluent tube assembly; the side wall of the upper cover assembly is connected with the upper part of the main body assembly, and the main body assembly is nested in the decorative cover; a reagent system in the kit comprises an RT-RAA reaction system and a CRISPR reaction system which are stored in the detection kit at normal temperature in the form of freeze-dried microspheres and closed liquid, and a lysis solution and a diluent are released through the lysis solution tube assembly and the diluent tube assembly and react in the main body assembly. According to the invention, the problems that the existing fluorescence detection operation process is complicated, the instrument is not portable and depends on professional analysis and the like are solved, and the rapid, portable and user-friendly detection on the virus is realized based on the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) detection system.

The invention discloses an RAA-CRISPR / Cas12a detection system of a circle virus avian origin type 1 and application of the RAA-CRISPR / Cas12a detection system, and relates to the technical field of microbiological detection. The detection system comprises an RAA amplification primer pair, crRNA, an FQ-ssDNA probe, an FB-ssDNA probe and an LbCas12a nuclease, and is characterized in that the detection system comprises an RAA amplification primer pair; the RAA amplification primer pair comprises an upstream primer of which the nucleotide sequence is shown as SEQ ID NO.5 and a downstream primer of which the nucleotide sequence is shown as SEQ ID NO.8; the nucleotide sequence of the crRNA is as shown in SEQ ID NO. 11. The RAA-CRISPR / Cas12a detection kit established by adopting the detection system developed by the invention has the advantages of high efficiency, high specificity and high sensitivity, can be used for on-site rapid detection of GyVg1, and provides a powerful diagnostic tool with a cost advantage for a basic-level working environment.

Provided herein are nucleic acids useful as guide nucleic acids (gNAs), e.g., guide ribonucleic acids (gRNAs), in a CRISPR system wherein the guide nucleic acids contain one or more modifications to one or more nucleotides, use of such guide nucleic acids in modifying cells, and other uses wherein CRISPR Cas proteins are utilized.

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 an intracellular mycobacterium tuberculosis in-situ nucleic acid detection method based on CRISPR / Cas and application, and the method comprises the following steps: S1, designing crRNA and a primer pair of a mycobacterium tuberculosis target gene sequence, and synthesizing a single-stranded DNA fluorescent probe; s2, preparing a standard solution containing a target gene sequence, and incubating with a CRISPR / Cas12a system; s3, extracting nucleic acids of different types of bacteria, uniformly mixing the nucleic acids of different types of bacteria with the CRISPR / Cas12a system, and incubating; s4, incubating an attenuated strain of mycobacterium tuberculosis and the DNA fluorescent probe, infecting macrophages, and incubating to obtain infected cells; s5, electrically transferring a CRISPR / Cas12a system into the infected cells in S4 by utilizing an electroporation technology, incubating, and detecting the intensity of a fluorescence signal; and S6, performing quantitative and positioning analysis on the detected fluorescence signal to complete detection, and directly introducing a CRISPR / Cas12a system into the macrophage by adopting an electroporation technology to realize intracellular in-situ detection, thereby avoiding the operation complexity of a nucleic acid extraction link and the loss of detection sensitivity in the prior art.

The invention belongs to the technical field of molecular biology, immunology and virology, and discloses an RT-LAMP-CRISPR / Cas12b-based micro-fluidic detection kit and method for beet western yellow virus, isothermal amplification LAMP and CRISPR / Cas12b detection are connected in a reaction chamber by utilizing a raffinose dynamic diffusion system, and simultaneous detection of multiple samples is realized through a micro-fluidic chip. The detection method effectively avoids aerosol pollution, has the advantages of excellent specificity, sensitivity and visualization, is simple and convenient to operate, does not need expensive instruments and equipment, can simply, conveniently and quickly detect whether pepper and beet samples are infected by beet western yellowing virus in the field, and improves the detection efficiency.

Systems and methods for targeted gene modification, targeted insertion, perturbation of gene transcripts, and nucleic acid editing. Novel nucleic acid targeting systems comprise components of CRISPR systems and non-LTR retrotransposon elements.

The invention discloses a kit for visually detecting an H5 subtype avian influenza virus based on an RPA-CRISPR / Cas12b system and application of the kit, and belongs to the technical field of molecular detection. The kit comprises an RT-RPA (Reverse Transcription-Recombinase Polymerase Amplification) primer pair and sgRNA1 The RT-RPA primer pair comprises a forward primer with a nucleotide sequence as shown in SEQ ID NO.1 and a reverse primer with a nucleotide sequence as shown in SEQ ID NO.2; and the nucleotide sequence of the sgRNA1 is as shown in SEQ ID NO. 7. According to the kit, nucleic acid amplification and detection are synchronously completed through a tubular reaction system, the detection sensitivity reaches 1 copy / mu L, the coincidence rate with fluorescent RT-PCR is 100%, and no cross reaction exists on H7, H9, NDV and IBV. The kit does not need a complex instrument, is suitable for rapid screening on a basic level site, and provides an efficient solution for prevention and control of avian influenza.

Disclosed are lipid nanoparticle (LNP) delivery systems that specifically target T cells. The LNP delivery system comprises antibodies conjugated to the surface of the LNP, e.g., via maleimide chemistry, that target at least two T cell surface proteins, e.g., CD3 and CD28. The LNP delivery system can have a single population of LNP conjugated to either a bispecific antiCD3 / antiCD28 antibody, or two monospecific antiCD3 and antiCD28 antibodies, or two populations of LNP wherein each population comprises a monospecific antibody. The payload of the LNP delivery system can be, e.g., mRNA encoding chimeric antigen receptors (CAR), a linear DNA fragment or a plasmid encoding chimeric antigen receptors (CAR) or therapeutic proteins such as antibodies, components of a gene editing systems (e.g., CRISPR-Cas), small molecules, antibody-drug conjugates (ADC), and any combination thereof, either encapsulated in the LNP or attached to its surface (e.g., conjugated). Also provided are lipids, pharmaceutical compositions, kits, and methods of treatment.

The invention discloses a method for detecting dengue virus based on RPA / RAA / ERA / MIRA-CRISPR / Cas12a, which comprises the following steps: extracting nucleic acid to obtain a sample to be detected, putting the extracted sample in a sterile and disinfected sample container, contacting the sample to be detected with a composition, a product combination, a reagent, a kit, a system or a system, determining the signal change after the sample to be detected is contacted with different substances, and determining the detection result of the dengue virus based on RPA / RAA / ERA / MIRA-CRISPR / Cas12a. In the same container, a to-be-detected sample is in contact with a composition, a product combination, a reagent, a kit, a system or a part used for RPA / RAA / ERA / MIRA nucleic acid amplification in the system, and then is in contact with a part used for CRISPR nucleic acid detection, the RPA / RAA / ERA / MIRA and CRISPR / Cas12a detection methods are combined, a rapid, simple, sensitive and specific dengue virus detection method is realized, the detection sensitivity is high, and the detection time is short. The reaction conditions of virus detection are further optimized.

The invention relates to the technical field of biosensing, in particular to a method for synergistically activating a CRISPR / Cas12a system based on a T-type structure and application of the method, the CRISPR / Cas12a system can be activated by utilizing two split DNA activation chains and crRNA to form the T-type structure, the activated CRISPR / Cas12a system has trans-cleavage activity, an FQ-ssDNA probe modified with an FAM fluorophore and a BHQ1 quenching group can be subjected to trans-cleavage, a fluorescence signal is recovered, and the fluorescence signal is recovered. Through reasonable design of sources of activation chains corresponding to different targets, detection targets can be expanded from traditional nucleic acid detection to non-nucleic acid targets such as proteins and metal ions, so that biosensing application of multiple targets is realized, in addition, design based on a T-type CRISPR / Cas12a synergistic activation strategy has great flexibility and universality, and the application prospect is wide. The method can be used together with various amplification strategies and detection platforms, and the sensitivity of the analysis method is effectively improved.

The present invention relates to an engineered Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system comprising engineered dual guide nucleic acids (e.g., RNAs) capable of activating a CRISPR-Associated (Cas) nuclease, such as a type V-A Cas nuclease. Also provided are methods of targeting, editing, and / or modifying a nucleic acid using the engineered CRISPR system, and compositions and cells comprising the engineered CRISPR system.

The invention discloses a detection system for simultaneously detecting circle virus poultry source type 1 and circle virus human source type 1 and application of the detection system, and relates to the technical field of biological detection. The detection system comprises an RAA amplification primer pair of GyVg1, an RAA amplification primer pair of GyH1, a 12-crRNA-3 (12-crRNA-3), a 13-crRNA-2 (13-crRNA-2), an ssDNA (single-stranded deoxyribonucleic acid) probe, an ssRNA (single-stranded ribonucleic acid) probe, an LbCas12a nuclease, an LbuCas13a nuclease and a T7 transcriptase. According to the present invention, based on GyVg1 and GyH1 conserved genes, the specific crRNA and the RAA primer are respectively designed, and the RAA amplification technology and the CRISPR-Cas fluorescence detection system are combined so as to establish the detection system for simultaneously detecting GyVg1 and GyH1; the detection system can detect two different subtypes of circle viruses at the same time, the detection efficiency is improved, and a new technical means is provided for dynamic research of virus infection.

The present invention provides a model or tool for predicting the editing efficiency of sgRNA in a CRISPR / Cas gene editing system, in particular a CRISPR / dCas epigenetic editing system, a training and prediction method thereof, and a related computer system, computer storage medium, and application. In particular, one or more sgRNA and target gene related epigenetic features are added to an input of the model.

The invention provides a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas-based kit for isothermal rapid visual detection of H1N1 influenza virus and mycoplasma pneumoniae and a detection method thereof. The kit and the detection method provided by the invention can be used for timely, sensitively and specifically detecting a nucleic acid target in a sample. The invention provides a lateral flow analysis (LFA) detection platform of a three-stage amplification system integrating recombinase polymerase amplification (RPA), CRISPR / Cas12a and catalytic hairpin assembly (CHA) circulation, the detection platform is used for sensitively and accurately detecting mycoplasma pneumoniae DNA (MP DNA) and influenza A virus subtype H1N1RNA (H1N1RNA), and compared with other researches based on an LFA-CRISPR / Cas12a system, the detection platform has the advantages that the detection efficiency is greatly improved, and the detection cost is reduced. According to the kit and the detection method provided by the invention, nucleic acids of two pathogens can be visually detected on a single LFA at the same time, and the sensitivity is high.

The invention discloses a novel tsRNA high-sensitivity detection kit based on CRISPR (clustered regularly interspaced short palindromic repeats) / Cas13a-Cas14a, and belongs to the field of tumor marker detection. The tsRNA (tRNA-derived small RNA) is a diagnostic marker which is found recently and has great potential. In order to realize the clinical transformation of tsRNA as soon as possible, the tsRNA detection kit is developed on the basis of nuclease P1 combined with CRISPR / Cas13a-Cas14a linkage dual systems, and comprehensive, ultra-sensitive, high-specificity, low-cost and rapid tsRNA detection is realized through multiple signal amplification. Compared with the traditional technology, the sensitivity of the method is improved by 105-107 times, 1% of tsRNA in the mixture can be detected, and the method has more advantages in clinical application. Therefore, a new scheme is provided for application of the tsRNA in clinical diagnosis of diseases, and meanwhile, a reliable technology is provided for mechanism research of the tsRNA in occurrence and development of the diseases.

The invention belongs to the technical field of living cell imaging, and discloses a living cell nucleic acid imaging system and method based on CRISPR (clustered regularly interspaced short palindromic repeats) and a nano antibody and application, the system comprises: a modified dCas protein expression vector containing n antigen polypeptide tags; the modified sgRNA and / or crRNA expression vector comprises a specific guide sequence; the fluorescent nano-antibody expression vector contains m fluorescent nano-antibody coding sequences which are connected in an operable manner, and fluorescent protein serving as an internal tag is inserted between nano-antibody splitting sites, can be combined with a polypeptide tag carried by dCas protein and can stably emit light; and the fluorescent nano antibody in a free state can be self-degraded, so that the fluorescent nano antibody does not emit fluorescence. According to the invention, the single-copy nucleic acid sequence, especially the imaging marker of the single-copy nucleic acid sequence in a living cell, can be visualized with single-molecule resolution and high signal-to-noise ratio.

The invention provides an RPA-CRISPR-Cas12a system-based detection reagent for the drug resistance of a chilo suppressalis organophosphorus insecticide, a detection method and application, and belongs to the technical field of molecular detection. The invention provides a detection reagent for chilo suppressalis organophosphorus insecticide resistance, the reagent comprises a specific RPA2 primer pair designed for chilo suppressalis acetylcholin esterase ace-1 gene A314S resistance mutation site and crRNA, a double screening mechanism of RPA amplification and CRISPR-Cas12a is combined, wild type and mutant type genes can be accurately distinguished, and the detection reagent can be used for detecting the insecticide resistance of the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis. The rapid, sensitive and visual nucleic acid detection of the chilo suppressalis ace-1 gene A314S resistance mutation site is realized, and a visual detection means which is high in specificity, simple and convenient to operate, economical and efficient is provided for field on-site determination of chilo suppressalis resistance populations.

The invention relates to a method of modulating the expression of an endogenous gene in a somatic cell derived from an induced pluripotent stem cell (iPSC), comprising differentiating an engineered iPSC capable of stable or inducible expression of a CRISPR-related protein into a somatic cell prior to introduction of a guide RNA (gRNA) sequence. The invention also relates to the engineered iPSC, cells obtained by the method and uses thereof.

The invention discloses a rhodosporidium toruloides strain with high homologous recombination efficiency as well as a construction method and application thereof, and belongs to the field of oil-producing yeast engineering bacteria. According to the rhodosporidium toruloides strain, the rhodosporidium toruloides strain NP11-SaCas9-1 is used as an original strain, and an RAD52 gene is over-expressed. The modified yeast engineering bacterium provided by the invention can realize high homologous recombination through CRISPR system mediation, the homologous recombination efficiency reaches 90-100%, a multi-gene editing platform can be constructed on the basis, and the modified yeast engineering bacterium can be applied to the fields of synthetic biology, metabolic engineering, genetic engineering and the like, so that the modified yeast engineering bacterium is simpler, quicker, more convenient and more accurate in the genetic modification process. And the homologous recombination activity of the rhodosporidium toruloides can be improved by expressing the homologous recombination related protein Rad52, and the genetic engineering strategy can be used for modifying other unconventional yeasts.

The present invention relates to a nuclease polypeptide, such as a CRISPR nuclease polypeptide derived from a reference nuclease, which may be nuclease A, nuclease K or nuclease M, comprising a RuvC nuclease domain and a HNH nuclease domain. Also provided herein are gene editing systems comprising such nuclease polypeptides and gene editing methods using the gene editing systems.