221results about "Microinjection based" patented technology

This invention belongs to the fields of locust biotechnology and gene editing technology, specifically relating to a method for preparing locusts. Crz Methods using gene mutants and their application in locust control. This invention discovers that inhibiting gene mutants in locusts... Crz Gene expression can be used to control locusts. Crz The gene sequence is shown in SEQ ID NO:3. This invention successfully established a homozygous Crz mutant strain of the migratory locust using gene editing technology. It was found that the albino migratory locusts obtained after Crz deletion were more easily preyed upon by natural enemies than the wild-type locusts, exhibited significantly lower levels of the aggregation pheromone 4VA, and showed weaker adaptability to low temperatures compared to the wild type. This provides a new strategy for the biological control of migratory locusts and can also serve as a reference for the biological control of other pests.

The invention belongs to the field of animal model construction, and particularly relates to application of an NFATC2IP gene knockout mouse model. Experiments prove that after the NFATC2IP gene is knocked out, the mouse has the characteristic changes of ovarian weight reduction, volume reduction, estrus cycle disorder, fertility reduction, AMH reduction, FSH increase and the like. It is indicated that the NFATC2IP gene is a key factor for maintaining ovarian reserve and endocrine balance, and an NFATC2IP gene knockout mouse can be used for revealing the occurrence mechanism of ovarian dysfunction and can also be used for screening of novel ovarian protection drugs, research of a hormone regulation mechanism and discovery and verification of biomarkers.

The invention discloses construction and application of a Primrose syndrome non-human animal model, and belongs to the technical field of animal models and disease research. According to the invention, through a CRISPR / Cas9 technology, a codon CAC for coding 596th histidine in a No.14 exon of a mouse Zbtb20 gene is subjected to site-directed mutagenesis into CGC for coding arginine, and a hybrid mouse model for simulating pathogenic point mutation (p.H596R) of human Primrose syndrome is constructed. The model can stably reproduce key clinical phenotypes of the Primrose syndrome, including overgrowth after adult, serum IGF-1 rise, memory dysfunction and anxiety behaviors, and shows abnormal hippocampal neuronal development and synaptic transfer related pathways and the like. The invention provides an important experimental tool for deeply revealing the pathological mechanism of Primrose syndrome, developing drug screening and treatment intervention research and the like.

The disclosure is directed to compositions that comprise a plasmid DNA construct having a DNA sequencing encoding a therapeutic protein, or a fragment thereof, in vivo, along with methods of generating and manufacturing the antibody or therapeutic protein, as well as methods for preventing and / or treating a disease in a patient.

The present invention delivers a substance such as a reagent into a cell. A tubular body coated with an electroconductive polymer for introducing a substance into a cell and / or recovering a substance from a cell. The use of a tubular body coated with an electroconductive polymer to introduce a substance into a cell and / or recover a substance from a cell. A device for introducing a substance into a cell and / or recovering a substance from a cell, the device being equipped with a substrate and a tubular body coated with an electroconductive polymer, and the tubular body being positioned on the substrate. A system for introducing a substance into a cell and / or recovering a substance from a cell, the system being equipped with the abovementioned device, a voltage supply unit for supplying a voltage, and, as needed, electrodes.

A method for producing a target protein, comprising: providing a host cell, a vector comprising a nucleic acid sequence encoding the target protein, and a transposase; transfecting the host cell using the vector and the transposase to obtain transfected cells; culturing the transfected cells to form a culture; and harvesting the culture to obtain the target protein from the harvested culture, wherein the cells of the culture are not passaged before the culture is harvested.

The invention belongs to the technical field of GPCR drug research and development, and discloses a ratio type cAMP fluorescent probe RaCamp and a construction method of a transgenic mouse, the probe RaCamp is formed by series fusion of G-Flamp2 fluorescent protein and mCherry fluorescent protein through a flexible linker, and the preparation method of the probe RaCamp comprises the following steps: S1, obtaining an mCherry fragment; s2, a G-Flamp2 linear carrier is obtained; s3, homologous recombination; s4, converting and screening; the construction method of the transgenic mouse comprises the following steps: T1, designing a carrier; t2, microinjection of fertilized eggs; t3, embryo transplantation and reproduction; the invention has the beneficial effects that mCherry is added as an internal reference protein on the premise of keeping the sensitivity of the original G-Flamp2 probe, and the selected connecting peptide (GGGGS) 2 enables the expression quantity of the two proteins in cells to be accurately controlled to be 1: 1, so that the problem of background interference in cell observation of in-vivo imaging is well solved. And the sequence is inserted into a Rosa26-LSL box, so that the construction of a Rosa26-LSL-RaCamp mouse is realized.

The present application relates to animals or cells having a myostatin gene with a 12 base pair deletion in the second exon. The application may also include compositions capable of engineering a 12 base pair deletion in the myostatin gene to construct animals or cells. The application also relates to the use of the compositions to increase muscle mass.

The invention provides a construction method and application of a humanized MECP2 gene mouse model, and relates to the field of gene engineering. The model animal with the humanized hMECP2 gene is successfully prepared, the humanized hMECP2 protein can be normally expressed in the body of the model, and the model can be used for MECP2 gene function research and screening and evaluation of human MECP2 targeted drugs and therapies. The animal model prepared by the invention can be used for rapidly establishing more different MECP2 mutation humanized disease mouse models, and is applied to drug screening, drug effect research, related nervous system disease and tumor treatment and the like aiming at human MECP2 target sites, so that the research and development process of new drugs is accelerated, the time and the cost are saved, and the drug development risk is reduced. And a powerful tool is provided for researching the functions of the MECP2 protein and screening drugs.

The application provides a method for constructing an animal model of dilated cardiomyopathy and application, belongs to the technical field of biological medicine and animal genetic model construction, provides a new zebrafish model of dilated cardiomyopathy, is constructed by specifically overexpressing a truncated dnajb6b gene long isoform fragment in zebrafish myocardial cells, the nucleotide sequence of the gene fragment is shown as SEQ ID NO. 1, and the method for constructing the zebrafish model of dilated cardiomyopathy and application are provided, and it is further found that 4-phenylbutyric acid is applied to the preparation of a drug for treating dilated cardiomyopathy.

Disclosed herein are peptide hydrogel-based formulations. In some embodiments, the peptide hydrogel-based formulations encapsulate antibodies. In some embodiments, the peptide hydrogel-based formulations prevent formation of high molecular weight protein species.

The application provides a construction method of a humanized keloid mouse model, comprising the following steps: BAC plasmid construction and preparation; superovulation of experimental mice and collection of zygotes; pronuclear microinjection of zygotes; post-injection embryo transplantation; genotype identification; breeding and genetic analysis; and verification of construction results of the humanized immune system by immunohistochemistry and immunofluorescence. In the application, peripheral blood mononuclear cells treated by sCD27 and skin around keloids (homologous cells and tissues) are transplanted into NSG-MHC-DKO immune-deficient mice with overexpression of CD70 genes, so that HLA rejection of different homologous immune cells and tissues can be avoided, the internal microenvironment of keloids and the interaction between the internal microenvironment and the immune system can be restored to the maximum extent, and the influence of immune factors on the occurrence and development of keloids can be realized in vitro. The application first discovers and verifies that the activation of the CD27-CD70 axis can promote the occurrence and development of keloids, and provides a suitable animal model for the research and development of anti-keloid drugs, especially immunotherapy.

Provided herein are methods, computer programs, and systems for automated microinjection, for example, automated Intracytoplasmic Sperm Injection (ICSI), wherein creating, a first dataset of an oocyte and a holding device and a second dataset of an injection pipette; detecting the oocyte and the holding device in the first dataset and the injection pipette; selecting the image of the first dataset and of the second dataset where an equatorial plane of the oocyte / holding device and of the injection pipette has an improved focusing parameter; selecting images of the first and second datasets and labeling the pixels associated with the oocyte and to the injection pipette; creating an injection trajectory for the injection pipette to perform the ICSI using detected morphological structures; detecting when the oocyte is rupturing and when the spermatozoa has been released from the injection pipette into the cytoplasm of oocyte.

The invention belongs to the technical field of transgenic and biomedical research tools, and particularly relates to a method for constructing a transgenic fish system based on a Tol2 transposon system, which comprises the following steps: constructing a myo6b-lhfpl5b-DsRed plasmid by using myo6b as a promoter, lhfpl5b as a target gene and DsRed as a red fluorescent protein tag through a microinjection method, and constructing a Tol2 transposon system-based transgenic fish system. Co-injecting the plasmid containing the Tol2 transposase mRNA and the Tol2 transposase mRNA into a fertilized egg of an AB zebra fish strain, and screening generation by generation to obtain a transgenic fish strain with stable inheritance MET channel gene lhfpl5b marked hair cells. The spatio-temporal dynamic visualization of the living whole fish to the lhfpl5b is realized, the deafness-causing molecular pathway is clarified, and an in-vivo platform is provided for drug screening and intervention strategies.

This invention discloses a method for constructing a fast-growing, sterile zebrafish model, belonging to the field of fish animal model preparation technology. This invention is the first to discover the role of the CK gene in regulating fish growth rate and fertility: fish with simultaneous knockout of ck1 and ck2 genes exhibit rapid growth and gonadal abortion. Using the method provided by this invention, a fast-growing, sterile zebrafish model was successfully constructed. These zebrafish exhibited gonadal dysplasia and sterility, but also rapid growth. Compared to wild-type zebrafish, the 4-month-old double mutant zebrafish showed significantly increased body length and weight. The disclosed gene function and zebrafish model construction method can be used for breeding fast-growing, sterile fish varieties and studying related mechanisms, which is of great significance for improving the economic benefits of aquaculture and protecting ecological security.

The present application provides a kind of brown planthopper RNA binding protein NlRBP gene inhibitor and application, the NlRBP gene inhibitor is the dsRNA capable of inhibiting brown planthopper NlRBP gene expression, the nucleotide sequence of the dsRNA is as shown in SEQ ID NO.3.The present application also provides the application of the above-mentioned brown planthopper RNA binding protein NlRBP gene inhibitor in the prevention and treatment of brown planthopper.The prevention and treatment of brown planthopper is to inhibit the survival rate of brown planthopper.The survival rate of brown planthopper is realized by inhibiting the expression of brown planthopper RNA binding protein NlRBP gene.The present application also provides the application of the above-mentioned dsRNA in preparation product.The present application provides a new prevention and treatment strategy and target for brown planthopper prevention and treatment, establishes the technology based on RNAi green control brown planthopper, provides foundation and reference for the precision, sustained pest control of brown planthopper, and has good application prospect.

The invention discloses a construction method of a glomerular IgA deposition mouse model, and relates to the technical field of animal model development and glomerular IgA dyeing. Comprising the following steps: obtaining Cas9 mRNA (messenger ribonucleic acid), guideRNA (ribonucleic acid) and donor DNA (deoxyribonucleic acid) in an in-vitro transcription manner; the method comprises the following steps: carrying out microinjection on Cas9 mRNA (messenger ribonucleic acid), gRNA (guide ribonucleic acid) and donor DNA (deoxyribonucleic acid) into fertilized eggs of a C57BL / 6J mouse; carrying out genotype identification on the F0 generation of birth mice through PCR (Polymerase Chain Reaction) amplification and sequencing; mating the F0-generation positive mouse with a wild type C57BL / 6J mouse, and breeding to obtain an F1-generation mouse; and spontaneous glomerular IgA deposition appears in 24 weeks after the F1 generation heterozygote mouse is born. According to the method, accurate mutation is introduced by utilizing a homologous recombination repair mechanism to construct a glomerular IgA deposition model, the efficiency is high, and the off-target risk is low.

This invention discloses a relapse-resistant cell line for hypofractionated colorectal cancer and its applications, relating to the field of cell biology. This relapse-resistant cell line for hypofractionated colorectal cancer was deposited on January 2, 2025, at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, with accession number CCTCC NO: C202533. This invention provides a novel biomaterial for studying radiotherapy-resistant colorectal cancer by constructing a hypofractionated colorectal cancer cell line. The cell line was screened in mice with healthy immune systems, exhibiting good physiological relevance, which is helpful for studying the influence of the immune microenvironment on tumor cell radioresistance and the remodeling effect of radioresistant cells on the immune microenvironment. This cell line can also monitor tumor recurrence after radiotherapy in real time using fluorescence imaging.

The invention discloses a litopenaeus vannamei TAUD gene, an SNP marker, a detection primer, a kit and application of the litopenaeus vannamei TAUD gene. The sequence of the SNP marker is as shown in SEQ ID NO.1, the SNP site is located at the 504th site from the 5'end of the SEQ ID NO.1, and the polymorphic form is T / C type; the SNP site can be used for simultaneously screening body length and body weight to breed litopenaeus vannamei, and a primer designed according to the sequence of SEQ ID NO.1 or a kit prepared from the primer can be used for detecting the SNP site. The TT genotype of the SNP marker Chr2414603628 provided by the invention is used for selecting parents, so that the body length growth speed of a breeding population can be increased, an innovative tool is provided for molecular breeding of litopenaeus vannamei, the breeding period is shortened, and industrial development and efficiency improvement are assisted.

The application relates to a method for constructing a congenital pure red cell anemia zebra fish model, which comprises the following steps: performing directional mutation on a bysl gene in a wild-type zebra fish embryo to obtain F0 generation zebra fish; mating the F0 generation zebra fish with wild-type zebra fish to screen positive heterozygote F1 generation zebra fish from born zebra fish; mating the positive heterozygote F1 generation zebra fish with wild-type zebra fish to screen positive heterozygote F2 generation zebra fish from born zebra fish; and mating the positive heterozygote F2 generation zebra fish to screen positive homozygote zebra fish from born zebra fish as a congenital pure red cell anemia zebra fish model. The model can simulate the symptoms of congenital pure red cell anemia, and can be used for mechanism research, drug evaluation and new drug screening of the disease.

The present application belongs to the technical field of biological medicine, and relates to an RNA intracellular delivery tool, a system and application thereof. The present application provides an RNA intracellular delivery tool, which comprises an extracellular retractable injection system and an RNA binding protein. In the delivery tool, the RNA binding protein is combined with a cargo RNA molecule to form a complex, the complex is loaded into the inside of the extracellular retractable injection system, and an RNA intracellular delivery system is formed. The delivery system punctures the cell in the form of retractable injection and releases the cargo RNA molecule into the cell, thereby realizing precise RNA intracellular delivery. The present application overcomes the problems of insufficient targeting, efficiency and safety of the existing RNA delivery technology, and provides a solution for RNA therapy.

A system, method, and apparatus for delivering genetic material to a shoot apical meristem. The delivery system may have a biolistic particle delivery assembly having a gas acceleration tube and a bombardment chamber. The bombardment chamber has a microcarrier launch assembly comprising an opening adapted to receive gas for acceleration of the microcarrier system and to direct gas to a target. The delivery system may have a source of pressurized carrier gas operatively connected to a first end of the gas acceleration tube; and an extender. The extender has a channel configured to fit through the opening of the microcarrier launch assembly. The length of the extender facilitates the penetration of the microcarrier system to an L2 layer of the shoot apical meristem, conferring germline transformation and / or modification.

This invention provides the application of the NlP21 protein and its encoding gene in regulating plant resistance to brown planthoppers, belonging to the field of genetic engineering technology. This invention provides the sequence of the NlP21 protein and its encoding gene in brown planthoppers. This gene is of great significance for regulating plant resistance to brown planthoppers and regulating the growth of brown planthoppers. Inhibiting the expression of the NlP21 gene in brown planthoppers can effectively reduce their survival ability, while overexpressing the NlP21 gene in plants can effectively improve plant resistance to brown planthoppers. The NlP21 protein and its encoding gene provided by this invention have excellent application prospects and significant application value for reducing pesticide use, maintaining ecological balance, and promoting sustainable development.

The invention belongs to the technical field of animal reproductive biology and cell engineering, and particularly relates to a fluorescence labeling expression system for visualizing an oocyte nucleus maturation process and application of the fluorescence labeling expression system. The fluorescence labeling expression system comprises an eukaryotic expression vector pVenus-LMNB1; the eukaryotic expression vector pVenus-LMNB1 comprises a nucleotide sequence for coding the fusion protein Venus-LMNB1. According to the invention, a nuclear fiber layer protein stably expressed on a nuclear membrane is selected as a research object, a eukaryotic expression vector pVenus-LMNB1 is constructed by using a molecular cloning technology, an LMNB1 gene containing a green fluorescent label Venus is overexpressed in a porcine oocyte by using a microinjection technology, fluorescent protein labeling of a structural protein on the nuclear membrane is utilized, and the expression vector pVenus-LMNB1 is constructed. The expression positioning condition of the LMNB1 in the maturation process of the porcine oocytes is observed, and a powerful tool is provided for development of in-vitro culture related researches of the porcine oocytes.

The invention relates to the field of gene vector construction, in particular to construction and application of a gene vector integrating fluorescent screening and self-deletion functions, the gene vector comprises a conditional knock-out (cKO) vector and a gene knock-in (KI) vector which are both integrated with a Dre-Rox mediated self-deletion module and a fluorescent screening module, the construction method comprises the following steps: carrying out HindIII / EcoRI double enzyme digestion on a pUC19 vector, carrying out homologous recombination amplification on a target fragment, carrying out connection transformation and screening verification to obtain a qualified vector; the vector is applied to preparation of cKO / KI gene modified mice, and efficient screening of positive individuals is realized through fluorescence preliminary screening, genotype identification and fluorescence quenching detection. The positive screening workload and cost can be reduced by 70% or above, non-target elements are accurately cut off, interference is avoided, time-space accurate regulation and control are achieved, carrier construction is easy and convenient, repeatability is high, and the method is suitable for gene function research, disease model construction and drug target verification.

The present application relates to the field of biotechnology, and more particularly to a method for knocking out runx2b gene of heterologous octoploid Changfeng crucian and application thereof. The present application designs gRNA-1 and gRNA-2 aiming at all the alleles of runx2b from two sets of parent sources of heterozygous gynogenetic crucian and Xingguo red common carp in Changfeng crucian; the mixture of two gRNAs synthesized by in vitro transcription and Cas9 protein is introduced into the animal pole of single-cell stage of Changfeng crucian fertilized eggs by microinjection, which can efficiently knock out all the 8 alleles of runx2b-A and runx2b-B. Any gRNA can guide the CRISPR / Cas9 system to simultaneously edit the 8 alleles; gRNA-1 achieves 97.5% of synchronous knockout rate, and gRNA-2 achieves 95% of synchronous knockout efficiency; when the double gRNA collaborative cutting strategy is used, the complete editing efficiency of the 8 functional copies reaches 100%.

To provide a model nematode of climacteric symptom.SOLUTION: A nematode model for climacteric symptoms, which satisfies at least one selected from the group consisting of enhanced expression of nlp-58 gene, a functional and / or expression-reducing mutation of nlp-58 gene, enhanced expression of tkr-1 gene, and a functional and / or expression-reducing mutation of tkr-1 gene.SELECTED DRAWING: None

The invention belongs to the technical field of animal model establishment, and particularly relates to a non-alcoholic fatty liver animal model and an establishment method, and the method comprises the following steps: constructing an ABHD17a gene knockout mouse model; and hybridizing the mouse with the knockout ApoE gene and the mouse with the knockout ABHD17a gene to obtain the mouse with the knockout ApoE gene and the knockout ABHD17a gene, and feeding the mouse with high fat for 16 weeks to obtain the non-alcoholic fatty liver animal model.

The present application relates to a kind of construction method of atherosclerosis humanized mouse model, belong to genetic engineering and genetic modification technical field.The present application is first in ApoE and Ldlr double gene knockout mouse background and obtains NOD-4G gene knockout mouse by knockouting Prkdc and Il2rg gene in mouse, humanized atherosclerosis model is constructed by injecting human umbilical cord blood CD133+stem cell and high-fat feed feeding.Through to atherosclerosis degree analysis, it is found that the aorta and aortic root of the humanized atherosclerosis model constructed have a large number of plaque formation, and contain human CD45 + Leukocyte.This atherosclerosis humanized mouse model provides better animal genetic model for the analysis of the role of human immune cells in the occurrence and development of atherosclerosis disease, and has important significance for the research of human disease and the development of new therapy.

The invention discloses an application of a substance taking Angpt18 as a target in preparation of a medicine for treating and delaying senescence, and the key regulation effect of an Angpt18 gene in the occurrence and progression process of normal senescence and senescence-related diseases is found for the first time through systematic and in-depth research; further experiments prove that by inhibiting or down-regulating the expression of the Angpt18 gene, the senescence process can be obviously delayed, and the expressions such as hypomnesia and behavioral ability decline in the senescence process can be effectively improved. Therefore, the Angpt18 gene can be used as an important intervention target for senescence and related diseases thereof, can be used for developing drugs with senescence delaying or treatment effects, can also be used as a biomarker for evaluating the senescence degree and diagnosing and prognosing the related diseases of senescence, and has wide application prospects and clinical transformation values.