670 results about "Zebrafish" patented technology

The invention relates to the technical field of gene knockout, and particularly discloses a method for breeding a tcf25 gene deletion type zebra fish through gene knockout. The method comprises the steps of through a CRISPR/Cas9 gene editing technology, designing an appropriate targeting gene locus on a tcf25 gene of the zebra fish, synthesizing in vitro to obtain specific sgRNA and Cas9-mRNA, microinjecting into a cell of the zebra fish, and after culturing an embryo for 60h, selecting the embryo for carrying out genotyping, so that the effectiveness of the selected locus is verified. The method provided by the invention is lower in off-target rate, removes the tcf25 gene through interference, is beneficial to further revealing the whole process of cardiac morphogenesis and a molecular mechanism regulating the process through researching functions through a genetics means, and is of great importance on understanding a cardiac disease pathology and researching and developing a new therapeutic schedule medically.

The invention provides a method, primer and plasmid for constructing an EPO gene knockout zebrafish animal model and a preparation method of the primer and the plasmid, and belongs to the field of biomedicine. The method includes the steps of 1) establishment of an EPO gene knockout zebrafish CRISPR oligomer sequence plasmid; and 2) establishment and culture of the EPO gene knockout zebrafish animal model on the basis of CRISPR gene knockout technology; wherein the step 1) comprises 1-1) design and synthesis of the primer aiming at an EPO exon 2 region target sequence and 1-2) synthesis of the Escherichia coli plasmid containing the EPO target sequence. The method for constructing the EPO gene knockout zebrafish animal model can be used to construct the EPO gene knockout zebrafish animal model, and facilitates further understanding and research of EPO.

The invention discloses a preparation method of a zebrafish notch1b gene mutant. The preparation method includes: determining positions of knock-out target spots of the notch1b gene; utilizing pUC19-gRNA scaffold plasmid as a template, and performing PCR (polymerase chain reaction) amplification with primers T7-notch1b-sfd and tracr rev; subjecting a PCR product to purification and in-vitro transcription to obtain gRNA (guide ribose nucleic acid); introducing the gRNA and Cas9mRNA into a cell-stage embryo of zebrafish, and culturing to obtain the notch1b gene mutant stable in inheritance. Thepreparation method has the advantages that by the aid of the CRISPR/Cas9 (clustered regularly interspersed short palindromic repeats, CRISPR/CRISPR-associated genes, cas gene) technology, and by meansof selecting a specific section of targeting domain, the notch1b gene in the zebrafish is knocked out, other genes are protected from being 'injured accidentally', the zebrafish without the Notch1b gene is formed, and great significance is achieved on study of Notch signal channels.

The present disclosure provides methods of producing enucleated cells by photoablation. Such enucleated cells may be used as recipient cells for Somatic Cell Nuclear Transfer and cloning. The nuclear donor and/or enucleated recipient cells may be any fish cells, such as zebrafish, koi, or medaka fish cells. Such methods may be used to efficiently produce transgenic fish including by way of example zebrafish, koi, and medaka fish.

Positional cloning has been carried out to identify the gene responsible for the hypochromic anemia of the zebrafish mutant weissherbst. The gene, ferroportin1, encodes a novel multiple-transmembrane domain protein, expressed in the yolk sac. Zebrafish ferroportin1 is required for the transport of iron from maternally-derived yolk stores to the circulation, and functions as an iron exporter when expressed in Xenopus oocytes. Human and mouse homologs of the ferroportin1 gene have been identified. The invention includes isolated polynucleotides, vectors and host cells comprising nucleotide sequences encoding Ferroportin1 proteins and variants thereof, including those having iron transport function. The invention also includes polypeptides encoded by ferroportin1 genes and variants of such polypeptides, and fusion polypeptides comprising a Ferroportin1 or a portion thereof. Methods to produce a Ferroportin1, methods to produce antibodies to a Ferroportin1 and methods to identify agents binding to a Ferroportin1, which can be inhibitors or enhancers of Ferroportin1 iron transport activity, are also described. Inhibitors of Ferroportin1 activity can be used in a therapy for hemochromatosis.

The invention discloses a new method for screening an anti-liver injury medicament by using a model organism zebra fish. The method comprises the following steps: continuously molding the zebra fish serving as a molding object for 3 days by using preferred carbon tetrachloride at the concentration of 1mmol/L to obtain the zebra fish of a liver injury model, and then evaluating the anti-liver injury activity of different receptor medicaments by using ALT and AST levels in the liver tissue of the zebra fish and the pathological change condition of the liver tissue of the zebra fish. Experimental results show that the new method for screening the anti-liver injury medicament by using the model organism zebra fish has high molding success rate, can simulate an in vivo environment to perform quick screening, and has the advantages of strong operability, accurate experimental results, little amount of required reacceptance medicament, strong repeatability, low cost, wide application range, capability of performing batch screening experiments and high work efficiency.

The invention includes a zebrafish assay for cardiac response.

The present invention is directed to a novel, target-blind approach to drug discovery. The concept is to model human phenotypes in a teleost, such as a zebrafish, and then screen compounds, e.g., small molecules, for their ability to alter the phenotype. Because the screen is performed with a whole vertebrate organism and uses a phenotype as the output, the need to first identify target genes is eliminated. This approach is powerful because a single screen can theoretically detect drugs affecting any target relevant to the phenotype being observed, even if those targets are not yet characterized.

Systems and methods for screening zebrafish comprising, a storage device for at least temporarily storing an image of a zebrafish to be screened; a zebrafish atlas; and an operating device that automatically screens the zebrafish at least in part by automatically comparing one or more anatomical features of the zebrafish to one or more standards.

The invention discloses a wnt16 gene deletion type zebra fish. Compared with the prior art, specific genes in the genome of a living body can be muted more efficiently and precisely; moreover, the manufacturing is simple, the cost is lower, multiple sites on target genes can be sheared at the same time, and any number of single gene can be muted. The wnt16 gene deletion type zebra fish can be applied to research related with gene and skeleton growth and other research for finding whether deletion of wnt16 gene is related with growth of other organs such as heart or not, and thus the wnt16 gene deletion type zebra fish has a good medical research value. At the same time, the growth period of zebra fishes without wnt16 gene is obviously shortened, and the wnt16 gene deletion type zebra fish also has a good commercial value therefore.

A method of expressing in vivo heart-specific fluorescence in transgenic line of zebrafish is developed, which provides a research model for studying heart-related gene functions and performing gene therapies in the future. The method comprises the following steps. Firstly, a plasmid is constructed. This plasmid construct includes the upstream regulatory region, the exon 1, the intron 1, and the exon 2 of cmlc2 gene, cDNA of GFP, wherein the cmlc2 gene and GFP cDNA form a cassette, and inverted terminal repeats from adeno-associated virus are flanked at both sides of this cassette. The plasmid construct is linearized and microinjected into one-celled zebrafish fertilized eggs. Lastly, the heart-specific fluorescent expressed zebrafish are selected and the germline-transmitting transgenic strain is generated.

The invention provides zebrafish models of acute myelogenous leukemia (AML), as well as methods of using these models to identify therapeutic agents for treating AML.

The invention belongs to the field of drug screening and particularly relates to a method of creating a zebra fish peristalsis model and screening prokinetic medicaments. The method comprises the following steps of: (1) selecting the zebra fish; (2) inducing the model and treating the compound; (3) quantitatively analyzing through a fluorescence microscope. A fluorescent dye is applied in the invention for the first time to research the gastrointestinal peristalsis conditions, the new research method is a great complement for the existing research models, and the peristalsis model has important value in high throughput screening of the prokinetic compounds due to the maneuverability, non-invasion, simplicity and quickness.

The invention relates to a building method of a zebra fish hyperlipidemia model and application of the animal model to hyperlipidemia disease research and lipid-lowering drug screening. The building method of the zebra fish hyperlipidemia model mainly includes the steps of zebra fish selection, feeding of zebra fish by yolk powder, histochemical staining or fluorescent staining, image analysis and/or microwell plate analysis and statistical analysis. The building method has the advantages of simplicity, convenience, rapidity, economy, high efficiency, high throughput and the like, and the model can be used for hyperlipidemia disease research and lipid-lowering drug screening. The building method and application of the zebra fish bacterial infection model are of great significance to acceleration of research and development on lipid-lowering drugs and improvement on treatment of patients suffering from hyperlipidemia.

The invention relates to the field of compound detection, in particular to a method for screening mitochondria targeted compounds by using zebra fish. The method mainly comprises the following steps of: zebra fish selection, compound treatment, dyeing treatment and quantitative/qualitative analysis, wherein the dyeing treatment is that the dyeing treatment is performed on the zebra fish, on whichthe compound treatment is performed, for 0.5 to 1 hour at the temperature of 28 DEG C by taking JC-1 or DASPEI as a staining agent. The method covers the shortage that mitochondrial damage cannot be detected in vivo in the prior art by using an in vivo experimental analysis method for screening the mitochondria targeted compounds by using the zebra fish, is convenient, swift, economic and practical, and has the characteristics of high flux and high specificity.

The invention relates to a zebra fish behavior induced and analytical method and system. The system comprises a visual stimulus device, a camera and an analytical system, wherein, the visual stimulus device can at least induce the eye movement reaction behavior and optomotor reaction behavior of zebra fish; the camera is used for shooting the behavior of the zebra fish, so as to acquire video data; the analytic system carries out behaviouristics analysis on the input video data so as to obtain the behavior data of the eye movement reaction and optomotor reaction of the zebra fish. The method and system provided by the invention can be used for analyzing the eye movement reaction and the optomotor reaction of the grown-up zebra fish as well as the eye movement reaction of the infant zebra fish.

The invention relates to a preparation method of a GRNa gene knockout zebrafish strain, which comprises the following steps: screening sgRNA of a target GRNa gene, constructing a eukaryotic expressionvector carrying Cas9 and sgRNA expression elements, screening a positive zebrafish founder, and screening first filial generation and second filial generation. The expression condition of the GRNa gene is identified and the obtained GRNa gene knockout zebrafish strain is proved to normally mate and lay eggs and can be stably inherited to filial generations after being mutually intercrossed for more than three generations of the GRNa gene knockout zebrafish strain. And genome, mRNA and protein levels prove that the GRNa gene is hardly expressed. The invention can be used as a tool for zfGRNa gene research. Related experiments such as HE staining and immunohistochemistry prove that the GRNa gene knockout zebrafish strain shows an obvious pathological phenotype and can be used for researching mechanisms of GRNa in diseases such as nervous system diseases and metabolic diseases and screening related drugs; the invention can also be applied to scientific research and economic culture of zebrafish, and has good commercial value.

The present invention relates to the use of cannabidiol (CBD) for the treatment of tumours associated with Tuberous Sclerosis Complex (TSC). In particular the CBD was able to decrease the number and size of marker cells, pS6, in a zebrafish model of TSC. This is5 suggestive of a disease modifying effect whereby treatment with CBD could result in the reduction or prevention of the benign tumours that occur in TSC patients. Preferably the CBD used is in the form of a highly purified extract of cannabis such that the CBD is present at greater than 98% of the total extract (w/w) and the other components of the extract are characterised. In particular the cannabinoid tetrahydrocannabinol (THC) has been substantially 10 removed, to a level of not more than 0.15% (w/w) and the propyl analogue of CBD, cannabidivarin, (CBDV) is present in amounts of up to 1%. Alternatively, the CBD may be a synthetically produced CBD. In use the CBD is given concomitantly with one or more other drugs used in the treatment of TSC. Such drugs may include rapamycin and/or everolimus.

The invention discloses bifidobacterium animalis NX-6 and application thereof in preparation of lipid-lowering and weight-losing medicines, and belongs to the technical field of microorganisms. The preservation number of the bifidobacterium animalis NX-6 disclosed by the invention is CGMCC (China General Microbiological Culture Collection Center) No. 20114. The uninactivated and inactivated fermentation supernate, bacterial suspension and cell disrupted supernate of novel bifidobacterium animalis NX-6 show good effects of slowing down weight and body length increase, improving body mass index(BMI) and reducing fat area percentage on zebrafish, and have a potential weight losing effect; triglyceride (TG) and total cholesterol (TC) in a zebrafish hyperlipidemia model body can be obviously reduced, and a good hypolipidemic effect is shown. The novel bifidobacterium animalis NX-6 disclosed by the invention has a huge potential application prospect in the aspect of preparing medicines fortreating and/or preventing obesity and hyperlipidemia.