683 results about "Gene knockout" patented technology

The invention belongs to the technical field of plant molecular biology, and discloses an application of a soybean GmSW10 gene in reducing soybean plant height, grain weight and grease content and / or increasing protein content. The CDS sequence of the soybean GmSW10 gene is as shown in SEQ ID NO: 1. Knockout of the soybean GmSW10 gene can directly influence the plant height, the grain weight, the oil content and the protein content of soybean seeds, after the soybean GmSW10 gene is knocked out, the weight of soybean seeds and hundred grains is remarkably reduced, the plant height is reduced, the protein content is increased, and the oil content is reduced, which indicates that the GmSW10 gene is a key gene for regulating and controlling the plant height, the grain weight, the oil content and the protein content of the soybean, and the GmSW10 gene can be used for preparing the soybean GmSW10 gene for regulating and controlling the plant height, the grain weight, the oil content and the protein content of the soybean. And a new genetic resource is provided for genetic improvement of soybeans.

The invention discloses application of protein TaTDM and related biological materials thereof, and belongs to the field of plant breeding. The amino acid sequence of the protein TaTDM is as shown in SEQ ID NO: 2 or SEQ ID NO: 4 or SEQ ID NO: 6. The TaTDM gene knockout homozygous mutant strains tatdmm1 and tatdmm2 are obtained by carrying out gene knockout on wild type wheat Fielder, and it is found that at the temperature of 25 DEG C, the total floret number of each spike of the Fielder, the tatdmm1 and the tatdmm2 and the survival rate under the water shortage treatment condition are remarkably higher than those of the wild type wheat Fielder. The protein TaTDM has an important application value in regulation and control of wheat ear type characters and stress tolerance, provides a high-quality germplasm resource, and is of great significance in cultivation of drought-enduring plant varieties and improvement of crop yield.

The invention provides a resveratrol production strain and a construction method and application thereof, according to the strain, tyrAfbr, tyrB, aroE, pntAB, ompF, aroGfbr and accA genes are up-regulated, a carbon storage regulation factor csrA gene is knocked out, the transcriptional level of a fabF gene is down-regulated through a BBaJ23113 promoter, a TAL gene from Rhodotorula glutinis, a 4CL gene from Arabidopsis thaliana and an STS gene from Vitis vinifera are heterologously expressed on a genome, and the strain carries a high-copy plasmid PET-28a-WAL at the same time; the strain takes glucose as a carbon source, resveratrol is efficiently and stably synthesized from the beginning by adopting a fermentation method, the production cost is low, and efficient production of resveratrol is realized.

The invention provides a human-derived recessive retinal degeneration disease organ-like model based on Prom1 gene knockout. Specifically, the invention provides a specific gRNA targeting a Prom1 gene exon 9, and the specific gRNA can be used for efficiently knocking out the Prom1 gene of the human embryonic stem cell. The invention also provides a Prom1 gene knockout stem cell line derived from the human embryo pluripotent stem cell H9 and a retinal organ disease model. The retina-like organ can provide a research model for clinical treatment drugs.

The invention discloses sgRNAs of a specific targeting grass carp RIOK3 gene and a construction method and application of an RIOK3 gene knockout cell line, and belongs to the technical field of gene engineering. The invention provides sgRNAs of a specific targeting grass carp RIOK3 gene and a sequence of a primer group, and also provides a method for constructing a large-fragment knockout cell line of the RIOK3 gene in combination with the sgRNAs, and application of the gene knockout cell line in research on an anti-virus infection mechanism of grass carp. The invention discloses the construction of the grass carp gene in-vitro research model by knocking out the antiviral negative regulatory factor RIOK3 of the fish cells by using the CRISPR-Cas9 technology for the first time, provides an in-vitro research material and a new way for the analysis of an antiviral immune regulation molecular mechanism of the grass carp, and particularly has important guiding significance for the research of resisting grass carp reovirus infection.

The invention relates to an STING gene knockout TIL capable of enhancing anti-tumor activity and a preparation method and application of the TIL, in particular to a genetically engineered immune cell, and an STING gene in the genetically engineered immune cell is silenced or down-regulated. The ferroptosis resistance of the immune cell provided by the invention is remarkably enhanced, the immune cell can be better infiltrated into a tumor microenvironment, and the immune cell has a remarkable anti-tumor effect.

The invention discloses a sugarcane ScPP2C49 gene and application thereof, and relates to the technical field of plant biology, the nucleotide sequence of the ScPP2C49 gene is as shown in SEQ ID No.1, and the amino acid sequence of protein coded by the ScPP2C49 gene is as shown in SEQ ID No.2. The gene is induced to express by drought and abscisic acid, and the encoded protein of the gene is positioned in a cell nucleus. Functional studies show that after overexpression of the ScPP2C49 gene in the plant, the drought resistance of the plant is negatively regulated through various mechanisms of inhibiting an abscisic acid signal channel, hindering stomatal closure under drought stress, weakening active oxygen scavenging ability, reducing photosynthetic efficiency, inhibiting root growth and the like. The invention also provides a recombinant overexpression vector containing the gene and a method for obtaining a transgenic plant with reduced drought resistance by using the vector through an agrobacterium-mediated method. According to the invention, a new gene resource is provided for deep analysis of a plant drought-resistant molecular mechanism, and an important target gene is provided for cultivation of high-stress-resistance crop varieties through a reverse genetics means (such as gene knockout).

The invention relates to the technical field of biology, in particular to application of a rice biological clock gene OsRVED in regulation and control of rice salt stress resistance, and provides application of a rice OsRVED gene as shown in SEQ ID NO.1, a rice OsRVED gene CDS sequence as shown in SEQ ID NO.2 and protein as shown in SEQ ID NO.3 in regulation and control of plant salt stress response. According to the invention, transgenic rice with OsRVED gene mutation is obtained through a CRISPR (clustered regularly interspaced short palindromic repeats) editing technology. After the transgenic rice OsRVED gene knockout mutant is subjected to 180mM NaCl salt stress simulation treatment, the phenotype is observed, the survival rate is counted, it is confirmed that the osrved mutant shows the salt tolerance phenotype compared with receptor rice Zhonghua No.11, and it is indicated that the OsRVED plays an important role in the process of responding to rice salt stress.

The invention belongs to the technical field of genetic engineering application and plant breeding, and particularly relates to application of a rice OsHGO gene to improvement of high-temperature tolerance of rice. The OsHGO gene in japonica rice Zhonghua 11 is knocked out through a CRISPR / Cas9 system, and a homozygous mutant plant with the OsHGO gene knocked out is obtained. The phenotypes of the wild type and mutant plants after high-temperature treatment are observed, and it is found that the wild type has more serious leaf wilt than the mutant plants. It is shown that after the OsHGO is knocked out, the influence of the high temperature on plant growth is relieved, and the OsHGO gene regulates and controls the tolerance of the rice to the high temperature. Through mutation of the gene, high-temperature-resistant rice can be cultivated.

The invention belongs to the technical field of genetic engineering, and relates to the field of wheat genetic engineering, in particular to a gene TaABCG42 for regulating and controlling cadmium accumulation of wheat as well as an encoding protein and application of the gene TaABCG42. According to the invention, a TaABCG42 gene is knocked out by using CRISPR / Cas9, a TaABCG42 gene overexpression plant is constructed at the same time, and a homozygous mutant is obtained by using a Hi-Tom sequencing technology and PCR (Polymerase Chain Reaction) successive generation screening. A wild type TaABCG42 homozygous overexpression and knockout plant treated with a 40 [mu] mol / L Cd solution for 5 days is subjected to analysis such as metal content determination, subcellular component metal determination, Cd chemical form determination and Cd ion flow determination, and the Cd absorption and transport approach of TaABCG42 is analyzed, so that the physiological and molecular mechanism of the TaABCG42 metal transporter for absorbing and transporting Cd is explored, and the application prospect is wide. The understanding on the wheat ABC metal transporter is enriched, and a theoretical basis is provided for cultivating high-quality wheat with low Cd accumulation.

The invention belongs to the technical field of fish breeding, and discloses a method for creating alkali-resistant grass carp new germplasm based on a ca12 gene editing technology and application. Carbonic anhydrase ca12 is used as a target gene, a CRISPR / Cas9 gene editing technology is used for carrying out gene editing on wild grass carp, the genotype of the grass carp is identified through sequencing, ca12 gene knockout F0-generation chimera and the corresponding wild type thereof are selected to carry out carbonate alkalinity stress experiments, and based on experimental results, it is known that the ca12 knockout chimera of the grass carp has high alkali resistance, and can be used for preparing the grass carp. The invention provides a novel breeding method and germplasm material for culturing fishes suitable for alkaline water culture, and has important industrial application value.

The application discloses an Escherichia coli probiotic with high beta-alanine yield and application thereof, and the recombinant genetically engineered bacterium takes a wild-type Escherichia coli probiotic as a starting strain and is subjected to one or more gene edits, including Trc promoter strengthening of a panD gene, an aspC gene, an sdhC gene and a ppc gene, overexpression of a panD gene from Bacillus subtilis, an aspB gene from Corynebacterium glutamicum, an aspA gene and a pyc gene from Corynebacterium glutamicum on a plasmid pTrc99a, and knockout of a cycA gene. 600 Compared with an existing reported Escherichia coli probiotic beta-alanine production strain, the OD of the recombinant genetically engineered bacterium is 4.4 times that of the existing reported strain, and the beta-alanine yield is 6 times that of the existing reported strain, so that the recombinant genetically engineered bacterium has great industrial application potential.

The invention discloses an SOD2 knockout cell line and application of the SOD2 knockout cell line in anti-poxvirus, and relates to the technical field of antiviral research. According to the invention, sgRNA of targeted superoxide dismutase 2 (SOD2) is designed, the sequence of the sgRNA is shown as SEQ ID NO.1-SEQ ID NO.2, then a stable SOD2 knockout human-derived non-small cell lung cancer A549 cell line is established by combining CRISPR / Cas9 gene editing with a lentiviral vector delivery technology, single-cell cloning is obtained through multiple rounds of puromycin screening, and the SOD2 gene knockout A549 cell strain is successfully constructed. Vaccinia virus Tian Tan strain infection shows that the number of plaques of SOD2 knockout cells is obviously greater than that of the plaques (about 2.3 times) of normal cell infection, and the plaques are relatively large, so that fine SOD2 has the effect of limiting intercellular transmission of poxvirus. The invention lays a foundation for research and preparation of antiviral drugs.

The present application relates to the technical field of genetic engineering, and in particular to a high-yield sophorolipid gene knockout engineering bacterium, a construction method thereof and an application, the engineering bacterium taking Bombus bornensis Candida as a starting strain, and using a gF-Hph-gR nucleotide sequence to replace a gene sequence seq2 in a sophorolipid synthesis path for regulating a lactonization reaction ， The gF-Hph-gR nucleotide sequence is SEQ ID No. 1, and the seq2 gene sequence is SEQ ID No. 2. The method provided by the present application uses fusion PCR to construct a seq2 gene knockout box with a resistance screening marker, and the shortest time required is 1 day, which provides a new way for efficiently constructing a sophorolipid production engineering strain. The high-yield sophorolipid gene knockout strain provided by the present application lays a foundation for large-scale industrial production of sophorolipid and meets market demand, and has a wide application prospect, economic value and social benefit.

Metastatic uveal melanomas are highly resistant to all existing treatments. To identify actionable vulnerabilities, the inventors conducted a CRISPR-Cas9 knockout screen using a library composed of chromatin remodelers. They revealed that the histone H3 methyltransferase SETDB1 plays a critical role in metastatic uveal melanoma cell proliferation and survival. Functionally, SETDB1 knockdown triggers decreased expression of genes related to replication and cell cycle and promotes growth arrest associated with increased markers for DNA damage and senescence entry. Using pre-clinical model, they further demonstrated that anti-SETDB1 therapy tumor growth in vivo. The inventors identify SETDB1 as a new relevant therapeutic target for the treatment of metastatic uveal melanomas. The present invention relates to a method for treating uveal melanoma in a subject in need thereof comprising a step of administering said subject with a therapeutically effective amount of SETDB1 inhibitor.

The invention belongs to the field of plant biotechnology and plant molecular breeding, and particularly relates to a brassica napus BnabZIP1 gene and application thereof in prevention and control of sclerotinia rot of colza. According to the invention, a gene editing technical vector is constructed aiming at sgRNA of the BnabZIP1 gene, agrobacterium tumefaciens is transformed, then brassica napus is transfected, and a homozygous mutant plant after the BnabZIP1 gene is knocked out has stronger sclerotiniose resistance. Results show that the BnabZIP1 gene can be used as a molecular marker or a genetic engineering improvement target of sclerotiniose resistance of rape, is used for screening and creating disease-resistant rape varieties, and has a good molecular breeding application prospect.

The invention discloses a screening and expression platform construction method for non-essential gene loci of corynebacterium glutamicum, and belongs to the technical field of biology. The center sites CGLRS02940, CGLRS03085, CGLRS08555, CGLRS09020 and CGLRS13560 of the corynebacterium glutamicum identified by the invention have the remarkable advantages that when gene knockout or knockin operation is carried out, the sites do not interfere with the normal physiological metabolism process of strains, and stable expression of exogenous genes can be guaranteed; meanwhile, the gene editing efficiency is relatively high, and an editing target with high universality and high reliability is provided for genetic engineering modification of corynebacterium glutamicum.

The application discloses application of an OSA3 gene, relates to the technical field of genetic engineering, and the nucleotide sequence of the OSA3 gene is shown as SEQ ID NO. 3. The application research proves that the OSA3 gene has the ability of regulating the deep root ratio of rice, and by using the ability, drought-resistant transgenic rice can be developed for arid regions. Meanwhile, the OSA3 gene knockout mutant osa3-1 / osa3-2 constructed by the application has the increased number of shallow root systems, and can be used for developing transgenic rice resistant to poor soil for poor soil regions, and enhancing the adaptability of rice germplasm to the environment.

The application discloses a method for creating a new strain of Carassius auratus gibelii with resistance to Cyprinid herpesvirus 2 (CyHV2) infection, and the method is characterized in that a gene editing technology is used to target knockout gsdf-a a gene and gsdf-b a gene, and an experimental animal infection model of a homozygous knockout strain of the gene obtained by the technology is established. gsdf Results of the experimental animal infection model show that gsdf the CyHV2 infection resistance of the CyHV2 gene knockout Carassius auratus gibelii is significantly enhanced, the histopathological damage of the Carassius auratus gibelii after being infected with the virus is reduced, the expression of a virus protein ORF47 in liver tissue of the Carassius auratus gibelii is reduced, and the transcription level of a virus gene orf46r is significantly reduced. The new strain of Carassius auratus gibelii created by the method can avoid exogenous gene pollution, and has the advantages that the CyHV2 infection resistance of the Carassius auratus gibelii is significantly enhanced.

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 a regulation mechanism research method of depleted precursor CD8T cells, and relates to the field of immunology and molecular biology. Comprising the following steps: detecting the expression level of the ARHGAP9 gene in a chronic virus infection model; constructing a T cell specific ARHGAP9 gene knockout animal model; the influence of ARHGAP9 deletion on the frequency, phenotype and function of the Tpex cell is analyzed; a single cell transcriptome sequencing technology reveals that ARHGAP9 regulates and controls a downstream signal channel of a Tpex cell. By constructing a chronic virus infection model, detecting the expression dynamic state of the ARHGAP9 gene, knocking out an animal model by utilizing T cell specificity ARHGAP9, and combining flow cytometry, qPCR, single cell transcriptome sequencing and other technologies, the invention discloses the effect of the ARHGAP9 as a novel immune checkpoint molecule, and provides a theoretical basis for developing Tpex cell targeting immunotherapy.

The present application relates to the technical field of plant genetic breeding, and particularly relates to a Brassica campestris fatty acyl reductase BcFAR2 gene, an expression protein thereof and application, a base sequence of the Brassica campestris fatty acyl reductase BcFAR2 gene is shown as SEQ ID NO. 1, and an amino acid sequence of the expression protein is shown as SEQ ID NO. 2, the Brassica campestris fatty acyl reductase BcFAR2 gene is cloned, and BcFAR2 gene overexpression Brassica campestris sterile plants can restore pollen activity; gene knockout Brassica campestris fertile plants can significantly reduce pollen activity; in Arabidopsis thaliana, overexpression of the gene can significantly improve salt tolerance of the transgenic plants; and in Brassica campestris, overexpression of the gene can significantly enhance salt tolerance.

A microbial gene editing scheme automatic design method, system and storage medium; the method comprises: selecting a design object for different types of strains, the design object is bacteria or fungi; after determining the target gene, obtaining the target gene information from the NCBI database; the design scheme of bacteria comprises the following steps: determining the gRNA design region; selecting the optimal gRNA sequence; doing sequence complexity and GC content analysis; the design scheme of fungi uses a frameshift scheme: the gRNA of the frameshift scheme is set at any position not overlapping with other genes after the start codon in the gene; select gRNA sequence in gRNA design region, select the optimal gRNA sequence; design two pairs of four primer sequences for homologous arm identification primer; this scheme can replace the complicated and time-consuming manual scheme, save a lot of time, solve the problem of high cost in the design of the best site and the design of sgRNA sequence scheme of artificial gene knockout.

The invention discloses a genetic engineering strain of high-yield secondary metabolite Fumagillin as well as a construction method and application of the genetic engineering strain, and the genetic engineering strain is obtained by taking aspergillus fumigatus as a chassis bacterium through hosA gene knockout or hosA catalytic active site mutation. According to the method, a hosA gene knockout strain (delta hosA) and catalytic active site mutation strains (HosAD133A, HosAH175A and HosAD210A) are constructed by means of molecular biology, and the mutant strains are subjected to liquid or solid fermentation under the culture condition of 37 DEG C, so that the yield of Fumagillin can be remarkably increased. According to the method disclosed by the invention, the inhibition on a Fumagillin biosynthetic pathway is relieved and the metabolic flux is enhanced by utilizing the activity change of histone deacetylase caused by hosA defects, and an efficient and directional production method for high-yield Fumagillin is provided, so that the method has an important industrial application value.

The invention discloses a method for efficiently producing general type CAR-T. The general type CAR-T. Specifically, the invention provides a method for efficiently producing universal CAR-T. The method comprises the following steps: (s1) providing a T cell to be edited, and carrying out activated culture on the T cell to obtain an activated T cell; (s2) gene introduction: transfecting the activated T cell by using the LVV integrated with the CAR gene so as to obtain the T cell of which the genome is integrated with the CAR gene; (s3) gene knockout: transfecting the T cell obtained in (s2) by using LNP (Leukocyte Natriuretic Peptide) coated with Cas9mRNA (Cas9 Messenger Ribonucleic Acid) and sgRNA (Single Guide Ribonucleic Acid) so as to obtain a T cell in which the TCR gene and the CD7 gene are knocked out; and (s4) culturing the T cells obtained in the step (s3) so as to obtain the universal CAR-T. According to the method, the UCAR-T production cost can be remarkably reduced, the preparation period is shortened, and commercialization of shelf-type cell therapy is promoted.

The invention relates to the technical field of plant genetic engineering, and discloses a strawberry stolon regulation gene ZFP6 and application thereof, and the nucleotide sequence of the gene is as shown in SEQ ID NO.1. Researches prove that the ZFP6 gene is specifically expressed in axillary meristem of strawberry in the adult-plant stage, and the GA20ox4 gene can be directly or indirectly activated to start a biosynthesis pathway of GA and drive axillary buds to develop towards stolons. Through genetic interaction analysis of the ZFP6 gene, the GA20ox4 gene and the RGA1 gene, the function of the ZFP6 gene as an axillary bud fate decision upstream regulation factor is further verified, the blank of an endogenous regulation mechanism of the strawberry stolons is filled, and the ZFP6 gene is proved to be a molecular target for specifically regulating formation of the strawberry stolons. The gene sequence, the primer, the gene knockout expression vector and the matched technical system provided by the invention lay a foundation for strawberry molecular breeding and industrial application.

The invention discloses application of an HIF1A inhibitor in preparation of a medicine for preventing and / or treating hypertrophic cardiomyopathy, and belongs to the field of biological medicine. For a rat model of spontaneous hypertrophic cardiomyopathy caused by MYH7B gene knockout, hypertrophic cardiomyopathy can be significantly relieved by intraperitoneal injection of the HIF1A inhibitor LW6, heart remodeling can be relieved, heart functions can be improved, and myocardial hypertrophy caused by silent MYH7B genes can be effectively resisted by using siRNA of mRNA transcribed by the HIF1A gene. The key mechanism of the HIF1A inhibitor for treating the hypertrophic cardiomyopathy is that the hypertrophic cardiomyopathy is prevented and treated by reducing the expression level of glycolysis key enzymes GLUT1, HK2 and the like, inhibiting the formation and accumulation of cardiac lactic acid and improving the ventricular remodeling of the hypertrophic cardiomyopathy. The results show that the HIF1A inhibitor can be used for treating and / or preventing the human hypertrophic cardiomyopathy and has the potential of being developed into the medicine for clinically preventing and treating the hypertrophic cardiomyopathy.

This invention discloses a method for inhibiting Tamdy virus infection and the use of TCP1 inhibitors. Specifically, this invention discloses a method for inhibiting Tamdy virus (TAMV) infection of host cells, the method comprising the step of reducing or inhibiting the expression level and / or activity of the TCP1 gene or its encoded protein in host cells, wherein the reduction or inhibition of the expression level and / or activity of the TCP1 gene or its encoded protein is achieved by methods selected from the group consisting of: gene mutation, gene knockout, gene interruption, RNA interference technology, gene editing technology, introduction of inhibitors of genes or proteins, or combinations thereof.

The present application relates to a gene editing vector of Caulobacter crescentus, a gene editing method thereof and a Caulobacter crescentus editing strain. The gene editing vector of the Caulobacter crescentus comprises the following gene elements: a plasmid backbone, a Pvan promoter, a spCas9M gene, a green fluorescent protein gene GFP, a gRNA, an upstream homologous arm and a downstream homologous arm of a target gene. The gene editing vector of the Caulobacter crescentus adopts a CRISPR / spCas9M-GFP gene editing system, constructs a high-efficiency, simple and scarless genome editing system in the Caulobacter crescentus, and can realize gene knockout and insertion.

The invention belongs to the technical field of biological medicines, and particularly relates to application of a PPARalpha agonist combined with an NPC1L1 inhibitor in tumor treatment. Specifically, it is proved for the first time that systemic PKM2 deficiency can cause increase of the total cholesterol level and promote growth of homologous allograft tumors, and the phenomenon is partially attributed to increase of the expression level of intestinal NPC1L1. In addition, in breast cancer cells, PKM2 gene knockout not only weakens the activity of a PPARalpha signal channel, but also improves the expression of NPC1L1. Fenofibrate and Ezetimibe can be used for cooperatively inhibiting proliferation and metastasis of tumor cells in vitro and in vivo, so that the Fenofibrate and Ezetimibe have important clinical significance and social value.