817results about "Microinjection based" patented technology

According to the brown planthopper lethal gene Nlpfam and the application thereof in biological prevention and control, the brown planthopper lethal gene Nlpfam can cause brown planthopper death after being interfered, so that dsRNA or an interference reagent is prepared through transcription of the brown planthopper lethal gene Nlpfam, the Nlpfam gene can be effectively interfered after the dsRNA or the interference reagent is injected into the brown planthopper, the abdomen of the brown planthopper becomes black, and the brown planthopper death can be inhibited. The brown planthopper can be effectively killed, and the biological control on the brown planthopper is realized.

The invention relates to the technical field of fish genetic breeding, in particular to a method for improving carassius auratus gibelio resistance to carassius auratus gibelio herpesvirus. A CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 (CRISPR associated protein 9) gene editing technology is utilized to specifically knock out the carassius auratus gibelio irf10-B gene to obtain a mutant genotype individual of which three irf10-B alleles are terminated in advance, namely the required carassius auratus gibelio with high resistance The invention also discloses two specific knockout targets of the irf10-B gene and an allele sequence corresponding to a mutant genotype individual in which three alleles of the irf10-B are terminated in advance. According to the invention, the function of the irf10-B in the prussian carp is verified for the first time, a new germplasm of which the capability of resisting the herpesvirus of the prussian carp is improved by 30% is rapidly obtained by knocking out the single gene of the irf10-B, and the new germplasm has important industrial value.

The invention discloses a method for targeted creation of a fast-growing new tilapia strain, and belongs to the technical field of fish cultivation. The method specifically comprises the following steps: (1) taking a fragment, as shown in SEQ ID NO.1, on a third exon of a pomc gene as a gRNA target sequence; (2) designing and synthesizing a gRNA primer sequence; (3) amplifying by adopting the gRNA primer sequence and the gRNA plasmid, and then transcribing and purifying to obtain a gRNA fragment; (4) transferring the gRNA fragments into fertilized eggs of tilapia mossambica through gene editing to obtain F0-generation positive fish; and (5) hybridizing the F0-generation positive male fish with the wild-type female fish, screening out F1-generation parent fish, and culturing the F1-generation parent fish to obtain F2-generation tilapia with homozygous deletion mutation. According to the invention, the pomc homozygous knockout line is established in the Nile tilapia mossambica for the first time, the new fast-growing line is a complete gene function deletion model, the cost is lower, the efficiency is higher, and the phenotype can be stably inherited.

The invention provides a full-autonomous cell operation system based on body intelligence, and relates to the field of cell operation, the system comprises a system hardware architecture and a system software architecture; the system hardware architecture comprises a micromanipulation execution system, a precision positioning system, a multi-mode sensing system and an intelligent calculation and control center. The micromanipulation execution system is used for cell environment interaction, the precision positioning system is used for sample bearing and environment control, the multi-modal sensing system is used for providing key information, and the intelligent calculation and control center is used for operation decision making; the system software architecture comprises a sensing layer, a planning layer, an execution layer and an interaction layer. According to the application, cell recognition, tool selection, path planning, operation execution and result feedback can be automatically completed without manual intervention; through tool autonomous switching and strategy dynamic adjustment, various operations such as cell clamping, injection, controllable deformation and micro-assembly can be completed, and the problems that traditional cell operation depends on manual work, efficiency is low, and consistency is poor are solved.

The invention discloses a preparation method and application of a self-reactive animal model. An autoreactive animal model is constructed based on an SWHEL system, it is found through the model that MTCH2 is a key target for regulating and controlling B cell autoreactivity, deletion of MTCH2 will cause generation of a large number of autoreactive B cells subjected to antibody category conversion, the disease risk is increased, and a basis is provided for research of B cell autoreactivity related mechanisms.

The invention discloses a construction method and application of an NMD escape model of an Alport syndrome mouse. A non-human animal model carrying Col4a5 gene c.4432delG frame shift mutation is prepared on the basis of a CRISPR / Cas9 gene editing technology. The method comprises the following steps: co-injecting gRNA of a 49 exon of a targeted Col4a5 gene, homologous recombinant donor oligonucleotide containing c.4432delG mutation and Cas9 nuclease into a mouse fertilized egg, and carrying out embryo transplantation to obtain an F0-generation mutant mouse; a mutation site is verified by combining PCR (Polymerase Chain Reaction) with sequencing, and a stably inherited mutation line is established through two generations of breeding. Through verification, the model accords with pathological characteristics of the Alport syndrome, can stably simulate typical clinical manifestation and pathological characteristics of the human X-linked Alport syndrome, and can be used as an important tool for research of the Alport syndrome.

The invention relates to the technical field of grass carp (Ctenopharyngodon idella), in particular to a method for knocking out a grass carp SMYD3 gene and application of the grass carp SMYD3 gene. The nucleotide sequence of the gRNA is as shown in SEQ ID NO: 1. The method comprises the following steps: synthesizing gRNA of a target gene through in-vitro transcription, then mixing the gRNA with Cas9 protein, introducing the mixture into an animal pole in a grass carp fertilized egg single-cell period by adopting a microinjection mode, and hatching to obtain the grass carp F0 generation with the target gene mutation. According to the application, the knockout of the grass carp gene is efficiently realized, and an effective technical means is provided for developing grass carp breeding and gene function research.

The invention provides an application of WNT7B in construction of a myopia animal model. Homologous genes wnt7ba and wnt7bb of the WNT7B in the zebra fish are knocked out or knocked down by applying a CRISPR / Cas9 gene editing technology, an animal model of which the eye axis length is remarkably increased, the eyeball movement frequency is remarkably reduced and the light response is reduced is obtained, and the result shows that the WNT7B is related to the high myopia. The model provides a favorable tool for pathogenesis and drug screening of high myopia, and has a good application prospect.

The invention discloses a recurrence-resistant cell strain for colorectal cancer large segmentation radiotherapy and application, and relates to the technical field of cell biology. The colorectal cancer large segmentation radiotherapy recurrence-resistant cell strain is preserved in the China Center for Type Culture Collection on January 2, 2025, the preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: C202533. By constructing the colorectal cancer large segmentation radiotherapy recurrence-resistant cell strain, a new biological material is provided for studying the radiotherapy recurrence-resistant colorectal cancer. The cell strain is screened in a mouse body with a sound immune system, has better physiological correlation, and is beneficial to research on the influence of an immune microenvironment on tumor cell radiotherapy resistance and the remodeling effect of radiotherapy resistant cells on the immune microenvironment. The cell strain can also monitor tumor recurrence after radiotherapy in real time through fluorescence imaging.

The invention discloses a construction method and application of a mouse model for specifically tracing heart valve cells, and belongs to the technical field of animal model construction. The model construction method comprises the following steps: (1) constructing a Wid1-Cre hybrid mouse before inserting a Cre recombinase coding gene into a termination codon of a mouse Wid1 gene; and (2) hybridizing the Wid1-Cre hybrid mouse with the Rosa26-tdTomato homozygous report mouse, so as to obtain the double transgenic mouse which simultaneously carries the Wid1-Cre gene and the Rosa26-tdTomato gene. According to the invention, on the basis of the expression characteristic that Wif1 is only limited to a valve area in the heart, Wif1-Cre is constructed; according to the Rosa26-tdTomato mouse model, specific tracing of heart valve cells is achieved, and an important experimental tool is provided for mechanism research of heart valve diseases and development of therapeutic drugs.

The invention relates to the technical field of fish genetic breeding, in particular to a method for improving grass carp reovirus resistance of grass carp. Four specific knockout targets of the herc4 gene are disclosed, the grass carp herc4 gene is specifically knocked out by utilizing a CRISPR / Cas9 gene editing technology, and a herc4 gene knockout individual, namely the high-resistance grass carp reovirus fish, is obtained. According to the invention, the function of the herc4 in the grass carp is verified for the first time, a new grass carp germplasm with improved grass carp reovirus resistance is rapidly obtained by knocking out the single gene of the herc4, and the herc4 has important industrial value.

The present application provides a brown planthopper lethal gene Nlpfam and its application in biological control. After the brown planthopper lethal gene Nlpfam is interfered with, it can cause the death of brown planthoppers. Therefore, by transcribing the brown planthopper lethal gene Nlpfam to prepare dsRNA or an interfering reagent, injecting it into brown planthoppers can effectively interfere with the Nlpfam gene, causing the abdomen of brown planthoppers to turn black, effectively killing brown planthoppers, and achieving biological control of brown planthoppers.

The invention relates to an application of a CTHRC1 inhibitor in preparation of a medicine for preventing and / or treating arthritis. Specifically, the invention provides application of an inhibitor or an antagonist of a CTHRC1 gene or a protein thereof, the inhibitor or the antagonist is used for preparing a composition or a preparation, and the composition or the preparation is used for (a) preventing and / or treating arthritis-related diseases; and / or (b) inhibiting cartilage cell senescence.

The invention provides a breeding method for constructing XX / XY sex determination all-female sterile fish and application. The method comprises the following steps: knocking out a catalytic enzyme coding gene of a fish sex steroid hormone synthesis pathway by using a gene editing technology to block the fish sex steroid hormone synthesis pathway, screening to obtain an effectively mutated homozygote XX sex genetic type pseudo male fish, and hybridizing the pseudo male fish with improved allotetraploid crucian and carp female fish to obtain the fish sex steroid hormone synthesis pathway. Therefore, an allotriploid population is obtained, and the population completely develops into female fishes and is sterile, so that the effect of obtaining all-female and sterile populations is achieved at the same time. The method provided by the invention has very strong applicability, and can be widely applied to aquaculture of fishes to obtain female unisexual groups of the fishes so as to improve the aquaculture yield, realize effective control on female fertility and protect ecological safety; the method has a wide application prospect in the fields of genetic breeding and ecological safety of aquaculture fishes.

The invention belongs to the technical field of gene engineering, and relates to a construction method of a mouse model capable of rapidly degrading MSTN protein, which comprises the following steps: sequentially inoculating a 5'homologous arm, 3xEAAAK, an FKBP1A gene synonymous mutation sequence, 3xEAAAK, a red fluorescent protein mCherry sequence and a 3 'homologous arm on a Donor vector to obtain a Donor recombinant vector; designing gRNA based on a third exon of the Mstn gene, mixing the gRNA, Cas9 protein and a Donor recombinant vector, injecting the mixture into a mouse fertilized egg, and performing in-vitro culture and transplantation to obtain an F0-generation mouse; and hybridizing the F0-generation mouse with a wild type, or continuously breeding the filial generation to obtain the knock-in mouse model. According to the invention, a dTAG targeted protein degradation technology is utilized, the obtained homozygous knock-in mouse is subjected to dTAG administration, and rapid and reversible degradation and removal of MSTN protein can be realized.

The invention provides a genome editor enDelIscB and a genome editor enDelIscB-T5E based on an IscB system, and further provides a base editor ICBE and a base editor IABE based on the enDelIscBD60A. According to the invention, a genome editor and a base editor which are more compact in size and higher in cleavage activity and insertion / deletion efficiency are realized by optimizing a single mutation site of the activity of a DelIscB protein cleavage plasmid, an optimized variant of the DelIscB protein and an optimized sgRNA variant of the DelIscB. According to the present invention, it is verified that the albinism model is efficiently constructed by using the enDelIscB editor and the enDelIscB-T5E editor through micro-injection of the mouse embryo, such that the enDelIscB editor and the enDelIscB-T5E editor have great potential in the field of genome editing.

The invention is applicable to the technical field of rabbit breeding, and provides a method for increasing the body weight of a rabbit based on a gene editing technology, which comprises the following steps: designing an action target sgRNA on a rabbit NR6A1 gene aiming at an NR6A1 p.P192S site; uniformly mixing the synthesized sgRNA and SpRY-CBE, injecting fertilized eggs, transplanting the fertilized eggs into fallopian tubes of receptor female rabbits, and feeding the receptor female rabbits until production; dNA is extracted from produced young rabbit ear tissues, PCR and sequencing are carried out, and if C at the fifth site in NR6A1 gene target sites is mutated into T, it is proved that single base mutation is obtained, and the NR6A1 gene edited rabbit is obtained. According to the invention, the rabbit variety with increased weight is successfully cultivated; a new thought of animal genetic breeding is developed by utilizing a gene editing technology, the method has application values in promoting improvement of genetic characters of rabbits and improving production performance and economic value of the rabbits, and powerful support is provided for high-quality development of the animal breeding industry in China.

The construction of a non-human primate Usher syndrome disease model having USH2A gene mutation, wherein a target sequence against the mRNA of the non-human primate USH2A gene is established and a corresponding sgRNA is synthesized, then a knockout vector is constructed, the sgRNA and a Cas9 mRNA are mixed and injected into a non-human primate embryo to obtain a non-human primate embryo having USH2A gene knockout, and then by means of embryo transfer, a non-human primate having USH2A gene knockout is obtained, which exhibits symptoms of Usher syndrome. Upon detection and identification of the knockout effect, the knockout of the USH2A gene can be effectively realized in a non-human primate cynomolgus monkey embryo and cynomolgus monkeys, which provides a new and effective disease model for the diagnosis and treatment of Usher syndrome in non-human primates.

The invention discloses a trpa1b gene mutant and a construction method of a tilapia low-temperature-resistant strain, and belongs to the technical field of biology. A tilapia mossambica model with a trpa1b gene knocked out is constructed based on a CRISPR / Cas9 gene editing technology, and the method comprises the following steps: determining a specific target sgRNA of the trpa1b gene, introducing the sgRNA and Cas9 protein with activity into a tilapia mossambica embryo through micro-injection, cultivating and screening F0-generation trpa1b knocked-out tilapia mossambica, and obtaining homozygote mutant tilapia mossambica through two-generation mating. The tilapia with the trpa1b gene knocked out shows higher low-temperature resistance, and particularly, the survival time of the tilapia under the low-temperature condition is prolonged. The invention provides an important model for low-temperature-resistant breeding of tilapia mossambica, and also provides a potential gene target for low-temperature-resistant strain breeding of other fishes in view of the conservative property of the function of the trpa1b gene.

This invention discloses a protein and gene related to resistance to neonicotinoid acetylcholine receptor competitive regulator insecticides in peach aphids, and their applications. It relates to the field of screening technology for neonicotinoid acetylcholine receptor competitive regulator insecticides in peach aphids. This invention obtains and clarifies an MpeABCC_4G protein and its encoding gene that is closely related to the resistance of peach aphids to neonicotinoid acetylcholine receptor competitive regulator insecticides. By knocking down the expression of MpeABCC_4G in resistant peach aphids using RNAi technology, the control efficacy of neonicotinoid acetylcholine receptor competitive regulator insecticides is significantly improved.

The invention discloses application of nilaparvata lugens NlGRP protein in prevention and treatment of nilaparvata lugens, and belongs to the technical field of prevention and treatment of nilaparvata lugens. By inhibiting the expression of the coding gene of the NlGRP protein in the brown planthopper, the growth and development of the brown planthopper can be obviously inhibited. Wide application prospects are realized; the method has great application value for reducing pesticide use, maintaining ecological balance and realizing sustainable development.

The invention provides a construction method and application of a dilated cardiomyopathy animal model, belongs to the technical field of biological medicine and animal genetic model construction, and provides a novel dilated cardiomyopathy zebrafish model which is constructed by specifically overexpressing a truncated dnajb6b gene long isomer fragment in zebrafish myocardial cells. The nucleotide sequence of the gene segment is as shown in SEQ ID NO. 1; the invention also provides a construction method and application of the dilated cardiomyopathy zebra fish model, and further discovers application of 4-phenylbutyric acid in preparation of drugs for treating dilated cardiomyopathy.

The invention provides a method for overspeed preparation of gene edited goldfish. The method comprises the following steps: S1, obtaining gobiocypris rarus without primordial germ cells as receptor fish; s2, designing a target spot for the goldfish amh gene, introducing a Cas9 / gRNA complex into a goldfish fertilized egg, performing sequencing verification to obtain a mutant individual, and feeding the mutant individual as a donor fish; s3, taking gonads of donor fish, cutting into pieces, performing enzymolysis digestion, filtering, and performing density gradient centrifugation to enrich donor germline stem cells; s4, transplanting the germline stem cells of the donor fish into the receptor fish body, and feeding to obtain the gene edited goldfish sperms and ova. S5, hybridizing and feeding goldfish amh mutant sperms and ova produced by gobiocypris rarus to obtain gene edited goldfish individuals. According to the invention, gobiocypris gobiocypris is used as a receptor, the cross-subfamily gene edited goldfish is used as a donor, and an efficient, stable and easy-to-popularize method for preparing the gene edited goldfish is provided by utilizing a belly borrowing reproduction technology.

The invention provides a construction method and application of a gene Idh1 conditional knockout mouse model and a preeclampsia heart injury animal model, and relates to the technical field of biology. According to the preeclampsia heart injury animal model, flox mice with conditional gene Idh1 knockout and mice with placental trophoblast specific expression Cre recombinase comate, PE-related phenotypes and typical symptoms of heart injury spontaneously appear in the double-positive pregnant mouse model, and the double-positive pregnant mouse model can be used for research of disease causes, diagnosis methods and drug research and development.

The present invention relates to a model mouse with a microglia-specific vitamin D binding protein (VDBP) gene knocked out and a construction method therefor. The construction method comprises the following steps: constructing a VDBPloxp / loxp gene mouse; hybridizing the VDBPloxp / loxp gene mouse with a Cre / ERT2 gene mouse to obtain a Cre+ / --VDBP+ / + gene mouse, wherein the microglia of the Cre / ERT2 gene mouse carry Cre gene; and inducing the expression of the Cre gene in the Cre+ / --VDBP+ / + gene mouse to obtain the model mouse with a microglia-specific VDBP gene knocked out. According to the construction method, a VDBP in the microglia of the model mouse can be specifically knocked out on the basis of Cre-loxP recombination, specific knockdown of the expression of the gene of the VDBP in the microglia is achieved, and the knockdown efficiency reaches 80% or more, facilitating more detailed study of the function of the molecule in the brain.

The invention belongs to the field of biological genetic engineering, and particularly relates to a method for constructing a congenital heart disease mouse model. The invention provides gRNA (guide Ribonucleic Acid) for targeting a mouse Tmed1 gene. Nucleotide sequences of the gRNA are as shown in SEQ ID NO: 1 and SEQ ID NO: 2. Meanwhile, the invention provides a method for knocking out the mouse Tmed1 gene based on a CRISPR / Cas9 technology to obtain the mouse with the congenital heart disease. The Tmed1 mouse model is successfully constructed, a reliable mouse model is provided for congenital heart disease and visceral ectopic syndrome patients, subsequent detection and verification of upstream and downstream target gene functions and subsequent verification of development of related target drugs are facilitated, and great convenience is brought to disease mechanism research and drug research and development verification work.

The invention discloses a method for constructing gene-free edited all-male fish based on a kdm6bb gene knock-down technology, which comprises the following steps: carrying out knock-down treatment on a fish kdm6bb gene by utilizing a CRISPR / Cas9 gene editing technology, and carrying out genotype identification to obtain a kdm6bb heterozygote (kdm6bb + / -); a kdm6bb heterozygous mutant capable of effectively reducing kdm6bb protein expression is obtained through selfing of heterozygous fish; pseudo female fishes with XY sex genetic types are screened through sex molecular markers, the pseudo female fishes are mated with wild XY male fishes, YY sex genetic super-male fishes without kdm6bb mutation are screened, and the YY sex genetic super-male fishes are hybridized with wild XX female fishes, so that all-male fish groups without kdm6bb mutation are obtained, unisexual culture of fishes is realized, sex control breeding efficiency is improved, and breeding cost is reduced. The present invention is widely applicable to aquatic economic fishes on the basis of XX / XY sex determination.

The invention discloses a method for constructing an Asiatic corn borer GBP gene homozygous mutant, which comprises the following steps: amplifying and cloning an Asiatic corn borer GBP gene sequence by using RACE technology, and the nucleotide sequence of the Asiatic corn borer GBP gene is as shown in SEQ ID No.1; two sgRNA target sequences are designed on a second exon of a GBP gene sequence of the ostrinia furnacalis, target sgRNA is synthesized through in-vitro transcription, the target sgRNA and Cas9 protein are fused in vitro to form an RNP mixture, and the RNP mixture is injected into ovum of the ostrinia furnacalis; the method comprises the following steps: carrying out PCR (Polymerase Chain Reaction) amplification on a target spot by using a target spot detection primer, sequencing to confirm a mutant individual, hybridizing and selfing to obtain a GBP homozygous mutant population. The Asiatic corn borer GBP gene homozygous mutant is constructed, so that the reproduction and immune functions of the mutant are remarkably reduced, and reference and research basis are provided for later development and formation of efficient pest biopesticide.

The invention discloses a construction method and application of a Prpf6 gene mutation induced single dose insufficient RP mouse model, and belongs to the technical field of molecular biology and biomedicine. The mouse model constructed by using the method provides a convenient, reliable and economic means for researching the relationship between PRPF6 gene mutation and retinitis pigmentosa diseases and the pathogenesis of the PRPF6 gene mutation, and provides an effective research model capable of stably inheriting in the research on the aspects of RP pathogenesis, treatment methods, drug screening and the like. The animal model is subjected to transcriptome and a series of phenotype detection, wherein a transcriptome result shows that Prpf6 gene knockout mouse retina has occurred a certain degree of metabolic abnormality, electrophysiological detection, fundus examination and Hamp; e staining results show that the shapes and functions of the retinal cell layer and the retina of the mouse are damaged, and the disease phenotype of RP is met.

The invention relates to the technical field of bioengineering, in particular to a coccinella septempunctata forkhead transcription factor Cs-FoxO gene as well as an inhibitor and application thereof. The invention provides a nucleotide sequence of a coccinella septempunctata forked head transcription factor Cs-FoxO gene as shown in SEQ ID NO. 1 and an amino acid sequence as shown in SEQ ID NO. 2; the invention also provides an application of the forkhead transcription factor Cs-FoxO gene inhibitor in relieving biological diapause. After dsRNA expression inhibition, the forked head transcription factor Cs-FoxO gene provided by the invention can relieve the diapause of the coccinella septempunctata, and the dsRNA of the Cs-FoxO gene can be used as a related preparation and product for relieving the reproductive diapause of the coccinella septempunctata, so that the development of the natural enemy industry is promoted, and the application prospect is wide.