123 results about "Microinjections" patented technology

A method has been developed to produce stable cell-matrix microspheres with up to 100% encapsulation efficiency and high cell viability, using matrix or biomaterial systems with poor shape and mechanical stability for applications including cell therapeutics via microinjection or surgical implantation, 3D culture for in vitro expansion without repeated cell splitting using enzymatic digestion or mechanical dissociation and for enhanced production of therapeutic biomolecules, and in vitro modeling for morphogenesis studies. The modified droplet generation method is simple and scalable and enables the production of cell-matrix microspheres when the matrix or biomaterial system used has low concentration, with slow phase transition, with poor shape and mechanical stability.

The invention discloses a construction method for a Sqstm1 whole-genome knockout mouse animal model and application. The mouse animal model is a mouse of which the Sqstm1 gene is knocked out. On the basis of a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 gene knockout technology, the Sqstm1 gene knockout mouse model is constructed. The construction method comprises the following steps: S1: designing sgRNA (Ribonucleic Acid) and Cas9 RNA, carrying out in vitro transcription to obtain mRNA, carrying out microinjection on the sgRNA and the Cas9 RNA with activity into amouse oosperm, to obtain the Sqtm1 gene knockout mouse; S2: identifying the Sqtm1 gene knockout mouse animal model. On the basis of the CRISPR/Cas9 gene knockout technology, the Sqstm1 gene knockout mouse animal model is constructed for the first time, and a convenient, reliable and economic animal model is provided for researching a relationship between Sqstm1 and diseases including autophagy, tumors and the like.

The invention relates to a continuous preparation method of superfine electrostatic-spinning fiber yarn, which comprises the following steps: (1) dissolving a polymer in a solvent to obtain a homogenous solution; (2) adding additives into the solution; (3) electrostatic spinning: feeding the spinning stock solution into a container, and extruding from a nozzle connected with a high-voltage anode under the control of a microinjection pump while controlling various electrostatic spinning parameters to keep a stable spinning state; (4) approaching a grounded needle tip to the nozzle to induce the electrostatic-spinning fibers to form bundles; (5) dragging the fiber bundles and twisting by a twisting machine; and (6) winding the twisted continuous yarns around a barrel by using a winding machine. The superfine electrostatic-spinning fiber yarn produced by the inventive method has wide application range.

The invention relates to a method for preparing a radiator element with a complicated shape, which belongs to the technical field of powder metallurgy technology. The method is characterized by comprising the steps: the mixture of metal or ceramic powder and organic matters is used as a raw material, and the metal or ceramic powder and the organic matter are sufficiently mixed according to a volume ratio from 0.45 to 3.8; mixing is carried out for the mixture at the temperature from 110 DEG C to 145 DEG C for 40 minutes to 90 minutes, and the group components of the organic matters and metal or ceramic powder and the organic matters are sufficiently mixed and pelletized to obtain evenly mixed injection feeding materials; a preforming blank of the radiator element with microstructure is obtained by adopting a powder microinjection forming technology; a degreasing step is carried out through the dissolution of the organic matters in the solvent and the heat dissolving mechanism in a heating process to eliminate the organic matters; and then, the radiator element with the complicated shape is obtained by sintering function. The method overcomes the defects of lower efficiency of machine processing technology, waste of the raw materials and organization aliquation caused by casting and processing, accurately controls the organization, increases the production efficiency and greatly reduces processing cost.

A preservation method for biological material having cell membranes includes microinjecting the cells with sugar; preparing the cells for storage; storing the biological material; and recovering the stored biological material from storage. The invention also features a method of culturing a cell in vitro using a hypertonic medium. Carbohydrate sugars such as trehalose, sucrose, fructose, dextran, and raffinose, may be used as bio-protective agents or in the culture medium.

The invention discloses a method for establishing an immunodeficiency mouse model. The method comprises the following steps of: (1) constructing DNA (deoxyribonucleic acid) which enables functions of an immune related gene in a mouse cell to be lost; (2) purifying mRNA (messenger ribonucleic acid), and cryopreserving in a super-purified water of RNAase free; (4) feeding a pregnant female mouse and a pseudopregnant female mouse under an SPF (specific pathogen free) level feeding condition; (5) taking a fallopian tube from the abdominal cavity of the pregnant female mouse in the step (4), and collecting fertilized ovum; (6) injecting the mRNA obtained in the step (3) into the pronucleus of the fertilized ovum in the step (5) by using a microinjection method; (7) transplanting the fertilized ovum obtained in the step (6) into the body of the pseudopregnant female mouse obtained in the step (4), and enabling the pseudopregnant female mouse to culture the second generation; (8) establishing a stable deficient mouse strain. The method has the advantages that a mouse model which completely does not have immunological rejection to allotransplantation can be obtained for testing.

A method of isolating target nucleic acid molecules from a solution comprising a mixture of different size nucleic acid molecules, in the presence or absence of other biomolecules, by selectively facilitating the adsorption of a particular species of nucleic acid molecule to the functional group-coated surface of magnetically responsive paramagnetic microparticles is disclosed. Separation is accomplished by manipulating the ionic strength and polyalkylene glycol concentration of the solution to selectively precipitate, and reversibly adsorb, the target species of nucleic acid molecule, characterized by a particular molecular size, to paramagnetic microparticles, the surfaces of which act as a bioaffinity adsorbent for the nucleic acids. The target nucleic acid is isolated from the starting mixture based on molecular size and through the removal of magnetic beads to which the target nucleic acid molecules have been adsorbed. The disclosed method provides a simple, robust and readily automatable means of nucleic acid isolation and purification which produces high quality nucleic acid molecules suitable for: capillary electrophoresis, nucleotide sequencing, reverse transcription cloning the transfection, transduction or microinjection of mammalian cells, gene therapy protocols, the in vitro synthesis of RNA probes, cDNA library construction and PCR amplification.

The invention relates to a paclitaxel-loaded sustained release nano-fibre material which comprises the components of bio-degradable polymer material and pure paclitaxel, the mol ratio of which is 1.33 to 10. The diameter of the nano-fibre is 90nm to 1.44micrometer, and the drug-loading rate can be regulated within the range of 0 percent to 100 percent. The preparation of the nano-fibre comprises the steps as follows: (1) the bio-degradable polymer material is solved in organic solvent and stirred to be solved completely to obtain lamella spinning solution A; (2) the white pure paclitaxel powder is solved in trifluoroethanol and stirred to be solved completely to obtain core spinning solution B; (3) clear transparent solutions A and B are respectively added into two injectors; the speed of a microinjection pump, the voltage of a static generator and the receiving distance between a grounded aluminum foil and a spinning needle are adjusted, and unordered drug-loaded nano-fibre is obtained by a coaxial electrostatic spinning technology. A nanometer control release system composed by the drug-loaded nano-fibre effectively controls the sustained release of the drug and is applied to the preparation of the drug for remedying malignant tumor.

The invention provides a method for constructing KARS-gene-mutated mouse models and application of the KARS-gene-mutated mouse models. The method for constructing the KARS-gene-mutated mouse models comprises the following steps: (1) carrying out target point construction: looking for a gene corresponding to nervous system diseases, and carrying out target point design; (2) designing Guide-RNA/Cas9mRNA, preparing plasmid, carrying out transcription, and then, injecting a transcription product into cytoplasm of a fertilized egg in a microinjection manner; (3) carrying out Founder mouse identification; (4) carrying out F1 mouse identification. According to the method for constructing the mouse models with the nervous system diseases and the application of the mouse models, provided by the invention, site-directed mutagenesis aiming at a KARS gene is carried out, so that a disease research on the model mice can be carried out under the condition of being closer to the occurrence and development of actual diseases, and a research result has a higher reference value.

The invention includes reagents and methods for the isolation of nucleic acids. The reagents described herein contain a nucleic acid precipitating agent and a solid phase carrier. The reagents can optionally be formulated to cause the lysis of a cell. These reagents can be used to isolate a target nucleic acid molecule from a cell or a solution containing a mixture of different size nucleic acid molecules. The disclosed reagents and methods provides a simple, robust and readily automatable means of nucleic acid isolation and purification which produces high quality nucleic acid molecules suitable for: capillary electrophoresis, nucleotide sequencing, reverse transcription cloning the transfection, transduction or microinjection of mammalian cells, gene therapy protocols, the in vitro synthesis of RNA probes, cDNA library construction and PCR amplification.

The present invention relates to a construction method of a human dcf1 gene transgenic drosophila melanogaster model. According to the invention, a drosophila melanogaster transgenic vector pUAST-dcf1 is constructed; a human dcf1 transgenic UAS drosophila melanogaster strain is obtained through microinjection; and human dcf1 transgenic drosophila melanogaster strains, with balance gene and genetic stability, of w, UAS-dcf1/Cyo and Tm6B/Tm2 are obtained through hybrid and backcross of the human dcf1 transgenic UAS drosophila melanogaster strain with Double Balance drosophila melanogasters of w, B1/Cyo and Tm6B/Tm2. The present invention utilizes microinjection gene engineering technology and drosophila melanogaster hybridization technology to construct the human dcf1 gene transgenic drosophila melanogaster model, and provides materials and new ideas for research on relative neurodegenerative diseases.

The invention discloses an oyster CRISPR/Cas9 gene editing method. The method comprises the following steps: screening a sgRNA target site sequence of a gene to be edited, designing a sgRNA sequence of a target sequence, and performing in vitro transcription to obtain a high-concentration target gene sgRNA; mixing sgRNA with Cas9 protein in equal volume, then reacting at 37 DEG C for 15 minutes, mixing with a phenol red solution in equal volume, introducing the mixture into oyster fertilized eggs by a microinjection method, and detecting oyster embryos or larvae after incubation for 6 hours toobtain corresponding gene mutants. According to the invention, an oyster gene editing technology is successfully established for the first time, and the technology can be used for studying the function of oyster genes and accurately modifying endogenous genes of oysters to provide a technical support for genetic improvement breeding of oysters.

A system for the administration of a bisphosphonate to a subject comprises a bisphosphonate formulation and a microinjection device. In an embodiment, the bisphosphonate formulation includes ibandronate sodium. In some cases, the bisphosphonate formulation includes an excipient. The bisphosphonate formulation may be delivered to the subject subcutaneously, transdermally or intradermally.

The invention discloses a new method for gene injection for a somatic cell nuclear transfer reconstructed embryo. The method comprises the following steps: target gene preparation, somatic cell nuclear transfer and a microinjection technology. An expression vector is constructed after a target gene is obtained, the cell nucleus of an oocyte is removed by virtue of micromanipulation, then a somatic cell is injected in the space of the denucleated oocyte, then an exogenous gene is directly injected in the somatic cell, the reconstructed embryo is constructed through one-step electric fusion and activation, and the embryo is transferred to obtain a transgenic progeny. According to the method disclosed by the invention, large-fragment cell gene transfection and screening are avoided, the transgenosis efficiency is increased, the time is saved, and the transgenosis application range is expanded. Importantly, the somatic cell is injected in a transpanent zone during a nuclear transfer operation process, thus microinjection for the exogenous target gene is facilitated.

The present invention discloses a method of preparing carbon nanotube by ethylene diamine double injection, which belongs to the carbon nanotube preparation technology. The method comprises the steps of: injecting ferrocene liquid hydrocarbon solution into a quartz tube reactor by an injector 1 driven by a microinjection pump, using ferrocene methanol solution and ethylene diamine as raw material, or ferrocene ethanol solution and ethylene diamine as raw material, or ferrocene toluene solution and ethylene diamine as raw material, or ferrocene dimethylbenzene solution and ethylene diamine as raw material, and using argon gas as carrier gas; and injecting ethylene diamine into a quartz tube reactor by an injector 2 driven by a microinjection pump; and obtaining the carbon nanotube by reacting for 40 min at 600~950 DEG C. The present invention has the advantages of that the insolubility and the chemical reaction between the different precursors can be avoided by using double injection to replace single injection; the structure of the carbon nanotube can be controlled by controlling the injection. The carbon nanotube of the present invention is adapted to functional material and composite material.

The invention relates to a construction method and application of a humanized mouse model, in particular to a construction method and application of a transgenic mouse model of a human Nbs1<c.657del5> gene. The method includes: constructing a 5bp deletion mutation-containing BAC carrier in the human Nbs1 gene, conducting pronuclei microinjection, and performing screening to obtain 3 stable transgenic lines for high expression and low expression of the human Nbs1 gene. The transgenic mouse involved in the invention has the phenotypes of delayed puberty, uniform shortening of body length and bone dysplasia at certain proportion in one of the lines, and a new mouse model is established for Nijmegen breakage syndrome diseases. At the same time, as the Nbs1 gene function impairment is closely related to cancers, the transgenic model can be applied to short-term carcinogenic tests in drug pre-clinical safety evaluation, thus providing a potential substitution model for traditional biennium carcinogenic tests and also providing an effective tool for research of carcinogenesis mechanisms.

The invention relates to a plant stress resistance enhancement related protein named as ZmHDZIV14. The protein is derived from a maize (Zea mays L.) inbred line B73, the amino acid sequence of the protein is represented as a sequence table SEQ ID NO:2, and an encoding gene sequence of the protein is a nucleotide sequence represented as SEQ ID NO:1. The invention further provides an application method of an encoding gene of the plant stress resistance enhancement related protein. According to the application method of the encoding gene of the plant stress resistance enhancement related protein, an expression vector carrying the ZmHDZIV14 gene is used for converting plant cells or tissue by the aid of conventional biological methods including application of Ti plasmids, Ri plasmids and plant virus vectors, direct DNA transformation, microinjection, electric conduction, agrobacterium tumefaciens mediated transformation and the like, a plant is cultured from the converted plant through tissue culture, and the plant with improved drought resistance and stress resistance is obtained. The expression vector carrying the ZmHDZIV14 gene is used for converting the plant cells or tissue by the aid of the conventional biological methods including application of Ti plasmids, Ri plasmids and plant virus vectors, direct DNA transformation, microinjection, electric conduction, agrobacterium tumefaciens mediated transformation and the like, the plant is cultured from the converted plant through tissue culture, and the plant with the improved stress resistance is obtained.

The invention discloses a method for constructing an anti-mammitis transgenic mouse model and a special vector for the method. An anti-mammitis-related protein composition comprises lysostaphin and peptidoglycan endolysin B30; the peptidoglycan endolysin B30 derives from a Streptococcus agalactiae phage B30, and an amino acid sequence of the peptidoglycan endolysin B30 is sequence 2 in a sequence table; and an amino acid sequence of the lysostaphin is sequence 3 in the sequence table. The experiment proves that a transgenic plasmid for mammary gland-specific expression of the lysostaphin and the peptidoglycan endolysin B30 is constructed by using a mammary gland-specific expression vector; and the anti-mammitis transgenic mouse model is constructed by a microinjection method, a transgenic mouse with an anti-bovine mastitis gene is obtained, and a technical model is provided for further producing anti-mammitis transgenic calves.

The invention relates to a method and device for preparing a nanofiber composite light-transmitting material, comprising: (1) preparing PA-6 spinning solution and PMMA spinning solution respectively, and spinning with a uniform blending device to obtain a composite nanofiber mat; ( 2) Lay the above-mentioned composite nanofiber felt in a mold, heat and pressurize, keep warm and pressurized for 20-60 minutes, take off the mold, and wait for complete cooling at room temperature to obtain the product. The device includes a receiving drum (1), a spinneret (2), a micro-injection pump (3) and a high-voltage power supply (4). The invention has the advantages of simple process, low cost and easy operation; the prepared composite light-transmitting material contains nanofiber reinforcement materials with a diameter less than 400nm, not only the original light transmission is not disturbed, but also the tensile mechanical properties can be significantly enhanced, thus It is very suitable for those occasions that have high requirements for light weight and mechanical properties.

The invention provides a preparation method and application of a transgenic mouse with severe immunodeficiency. The method comprises the step of knocking out a gene for encoding a gamma chain of an interleukin-2 receptor in a mouse chromosome through a CRISPR/CAS9 technology, exons 1 and 2 of a gene of a gamma chain of an interleukin-2 receptor are selected as knockout sites, and Cas9 messenger RNA and guide RNA are jointly injected into fertilized eggs of a mouse through microinjection. The mouse with severe immunodeficiency cannot generate T, B and NK immune cells, so that the greatest convenient condition is created for transplantation of human hematopoietic stem cells.

The invention discloses a method for breeding Macropodus opercularis with a transferred green fluorescent protein gene by using piggyBac transposons, which comprises the following steps of: firstly, constructing piggyBac transposons with green fluorescent protein genes and a helper plasmid capable of providing transposase; and secondly, transferring the two transposons into a germ cell of the Macropodus opercularis by using microinjection technology, and according to the transposons characteristics of the piggyBac transposons, introducing the green fluorescent protein genes into a genome of the Macropodus opercularis so as to make the gene inherited and expressed stably. An individual of transgenic Macropodus opercularis can be screened by means of the fluorescence-labeled gene, and the Macropodus opercularis with the transferred green fluorescent protein gene is more worthy of appreciation and represents a new variety of Macropodus opercularis with specific function. In addition, it is the first time for the piggyBac transposons to be used in researches on transgenic fish, and thus, the research space for transgenic aquatic animals is widened.

The invention provides a multi-degree-of-freedom electric micro-injection instrument. In the multi-degree-of-freedom electric micro-injection instrument, an X-axis direction sliding rail is arranged on a pedestal; an X-axis direction translation platform is arranged on the X-axis direction sliding rail; a micrometer head knob is arranged on one side of the X-axis direction translation platform; aY-axis direction sliding rail is arranged on the X-axis direction translation platform; a Y-axis direction translation platform is arranged on the Y-axis direction sliding rail; a micrometer head knob is arranged on one side of the Y-axis direction translation platform; a Z-axis direction sliding block sleeve is arranged on the Y-axis direction translation platform; a micrometer head knob is arranged on the top of the sleeve and is connected with a Z-axis direction sliding block in the sleeve; the middle part of the sleeve is provided with an opening through which a substrate is connected with the Z-axis direction sliding block; a connecting plate is arranged on the bottom side of the substrate and is connected with the Z-axis direction sliding block through a pitch adjusting nut; a microinjector and a motor are arranged on the substrate; one end of a screw rod is connected with a needle core of the microinjector; and the other end of the screw rod is connected with the output end of the motor. The multi-degree-of-freedom electric micro-injection instrument has a simple structure and the effects of stable and high-precision needle tip positioning and slow controlled injection.

The invention provides a method for constructing a mouse model with the autistic spectrum disorder. The method is characterized by comprising the step of injecting a methylation carrier and a gRNA carrier targeting an MeCP2 gene TSS region into cytoplasm by virtue of an embryo microinjection technique in the oosperm period of a mouse, so as to obtain the mouse model with the autistic spectrum disorder, wherein the methylation carrier contains a dCas9 fragment, a human source DNMT3L and a DNMT3A catalytic function domain. By virtue of the fixed point methylation carrier, the specific methylation of loci in cells can be effectively realized.

The invention relates to a method for preparing CNPase mutant zebra fish, and confirms that a mutant model can spontaneously generate bradycardia and even heart failure accompanied by severe heart failure. According to the invention, a CRISPR-Cas9 technology is combined with a microinjection technology, CNPase is specifically knocked out, a CNPase mutant F1 generation is prepared, and confirmation is performed through a Sange sequencing method; and through hybridizing and screening, a zebra fish homozygous mutant strain with heart specificity marked with green fluorescence is prepared, and it is confirmed that the CNPase homozygous mutant zebra fish can spontaneously generate cardiac hypertrophy and partially develop to heart failure through cardiology and kinetics indexes. The CNPase mutant zebra fish is prepared, an animal model is provided for researching functions of CNPase, and particularly great convenience is provided for researching related diseases of bradycardia accompanied by severe heart failure.