1015 results about "Gene knockout" patented technology

The invention relates to the field of rice gene engineering, and particularly provides OsSULTR2; the invention also discloses application of the 2 protein and the coding gene thereof in regulating and controlling the salt tolerance of rice. The protein meets the following conditions: B1) a protein with an amino acid sequence of SEQ ID NO.1; and B2) a fusion protein with the same function obtained by connecting a tag to the N end and / or C end of B1). The method is used for detecting OsSULTR2; the salt stress phenotype identification in the seedling stage is carried out on the transgenic rice with the gene knockout 1, 2, and the result shows that when the gene segment is deleted, the salt stress tolerance of the rice is improved, and the function and the application way of the gene are proved. Therefore, the OsSULTR2 of the present invention; the 2 protein and the coding gene thereof can regulate and control the salt tolerance of rice, and have important significance for cultivating salt-tolerant transgenic rice.

The invention discloses an application of an HERP gene knockout mouse model in obesity and metabolic disorder diseases. The invention finds that compared with a wild type high-fat group, HERP gene knockout can cause phenotypes such as early obesity, more serious obesity degree, increase of fat weight ratio, aggravation of adipose tissue inflammation, dyslipidemia and glucose metabolism disorder of mice during high-fat feeding. And the HERP gene is knocked out, so that the adipocyte differentiation is increased. The HERP gene knockout mouse is more sensitive to nutrition, obesity can occur more quickly during high-fat feeding, symptoms such as fat amplification, fat inflammation and glucose and lipid metabolism disorder are more serious, and the HERP gene knockout mouse can be used for molecular mechanism research and drug screening of obesity, fat tissue and glucose and lipid metabolism disorder. It is further found that cilostazol can significantly relieve mouse obesity and lipid metabolism disorder caused by HERP gene defects, but has no obvious weight losing effect on normal genotype mice, and a new strategy is provided for precise treatment.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a wheat magnesium transporter TaNIPA8-6B and application of a coding gene thereof to regulation and control of plant stripe rust resistance. Functional analysis is carried out on the wheat magnesium transporter TaNIPA8-6B by means of gene editing and overexpression, and it is found that the wheat magnesium transporter TaNIPA8-6B induces expression in both affinity and non-affinity system reactions of wheat and puccinia striiformis, so that a mutant plant of the gene editing and overexpression TaNIPA8-6B gene is created; according to the TaNIPA8-6B gene, the gene knockout mutant of the TaNIPA8-6B gene is identified to show susceptibility to stripe rust non-compatible races CYR23, the over-expression mutant plant shows disease resistance to stripe rust compatible races CYR32, the fact that the TaNIPA8-6B gene plays a positive regulation role in the disease resistance of wheat to stripe rust is clear, and a high-quality germplasm resource is provided for stripe rust resistance of wheat.

The invention relates to an L-valine production strain as well as a construction method and application thereof, the strain is obtained by transformation through a metabolic synthesis path of L-valine, and the construction method comprises the following steps: by taking Cornebacterium glutamicum ATCC 13032 (C.glutamicum) as an original strain, carrying out editing transformation such as gene knockout, gene overexpression, gene heterologous expression, gene double-copy expression and the like on a genome of the original strain by utilizing a metabolic engineering means. The L-valine production strain disclosed by the invention can be used for synthesizing L-valine from the beginning by taking glucose as a carbon source, has the advantages of high production rate, short fermentation period, high strain stability and low cost, and lays a foundation for realizing large-scale production of L-valine. According to the fermentation tank fermentation method, the L-valine is produced in a mechanical stirring type fermentation tank by using the L-valine production strain Va1-09, 86.4 g / L of L-valine is produced by fermenting for 36 hours, and the method has a good industrial prospect.

The invention belongs to the technical field of biology, and particularly relates to an SLC44A1 gene and / or SLC44A5 gene knockout animal model as well as a construction method and application thereof. The construction method of the animal model comprises the following steps: deleting exon regions from the fourth exon region to the thirteenth exon region of the SLC44A1 gene and / or the second exon region of the SLC44A5 gene in mouse brain oligodendroglia cells in a high-efficiency manner by utilizing gene editing and a Cre-loxP recombinase system, so as to realize the knockout of a target gene. The animal model constructed by the invention can be used for researching the regulation mechanism of the choline transporters (SLC44A1 and SLC44A5) on myelin sheath formation, deeply knowing the biological functions of the choline transporters (SLC44A1 and SLC44A5), simulating the occurrence and development processes of human related diseases, revealing the pathogenesis of the related diseases, providing a new thought for prevention and treatment of brain myelin sheath dysplasia, and providing a new foundation for the prevention and treatment of brain myelin sheath dysplasia. And an effective drug or a treatment mode aiming at the disease is screened by utilizing a gene knockout animal model, so that a new choice is provided for clinical treatment of the disease.

Disclosed herein are an engineered immune cell with CD7 gene knock-out and use thereof. According to the present invention, an sgRNA specifically targeting CD7 gene is designed and synthesized, which can accurately target CD7 gene to achieve gene knock-out with high knock-out efficiency. The provided sgRNA can be used for preparing a CD7-targeting engineered immune cell, and can be further used for preparing a CD7-targeting universal CAR-T cell

The invention discloses a pichia pastoris engineering bacterium for synthesizing bakuchiol from de novo by converting methanol as well as construction and application of the pichia pastoris engineering bacterium. A heterologous p-coumaric acid synthesis pathway is introduced to neutral sites of different chromosomes of pichia pastoris, a high-yield p-coumaric acid strain is obtained through gene knockout, overexpression or heterologous expression of tyrosine and phenylalanine synthesis pathway key genes of a shikimic acid pathway, a heterologous bakuchiol biosynthesis pathway is introduced on the basis, and a high-yield p-coumaric acid strain is obtained. After overexpression of bakuchiol synthetase, endogenous overexpression or heterologous expression of MVA pathway key genes and exogenous acetyl coenzyme A supply pathway genes, methanol concentration optimization and bacterial strain His4 gene supplementation, the yield of bakuchiol is effectively increased to 91.2 mg / L and is increased by 59.8 times compared with that of an initial bacterial strain, and the yield of bakuchiol in a 15L fermentation tank reaches 692.8 mg / L. The method has the characteristics of high conversion efficiency, low production cost, convenience in preparation, wide industrial application prospect and the like.

The invention discloses application of a rice transcription factor OsWRKY39 and a coding gene thereof in rice blast resistance of rice, and belongs to the technical field of gene engineering. According to the invention, an overexpression vector and a gene knockout vector of the rice OsWRKY39 gene are constructed, the vector is transferred into a Kitaake receptor material, an OsWRKY39 overexpression plant and an OsWRKY39 knockout plant are obtained, and the obtained transgenic plants are subjected to disease resistance analysis. Results show that knockout of the OsWRKY39 gene significantly improves the resistance level of rice to rice blast, which indicates that the OsWRKY39 negatively regulates the disease resistance of plants, and the encoding gene OsWRKY39 can be used as a target gene for improving the disease resistance of plants, has a good application prospect in cultivation of disease-resistant rice varieties, and lays an important foundation for rice resistance breeding.

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 invention relates to the technical field of plant genetic breeding, in particular to a rice sheath blight resistance related gene and an encoding protein and application thereof. The invention provides a rice sheath blight resistance related gene as well as an encoding protein and application thereof, the gene is OsSBR4 or a related gene thereof, the resistance of rice to sheath blight can be remarkably enhanced by knocking out the gene, and the resistance of rice to sheath blight can be reduced by overexpressing the gene. After agronomic traits of a transgenic line of the gene are investigated, it is found that the plant height, the ear length, the ear number, the thousand seed weight and the like are not affected, and it is indicated that the gene has important application value in sheath blight resistance molecular breeding. According to the invention, a novel rice sheath blight resistance related gene is screened and excavated, and the identification of the sheath blight resistance gene can provide a theory and a basis for breeding a sheath blight resistance rice variety.

The invention relates to the technical field of life science, in particular to a method for constructing an atrophic gastritis-based SPEM lesion model, which comprises the following steps: S1, constructing an Slc7a11 conditional knockout mouse strain: carrying out cage hybridization on an Slc7a11 flox / flox mouse and an ATP4b-Cre transgenic mouse, and knocking out a gastric wall cell specific Slc7a11 gene; s2, carrying out genotyping identification; the identification method based on the atrophic gastritis SPEM lesion model and the construction method based on the atrophic gastritis SPEM lesion model comprise the following steps: S1, pathological model verification: taking a mouse gastric mucosa tissue slice, quantitatively analyzing the proportion of an SPEM region by adopting an immunofluorescence double-standard technology, and confirming Slc7a11 knockout induced SPEM pathological characteristics in combination with the existence of SPEM in the gastric mucosa; and S2, long-term pathology evolution monitoring. Through gastric wall cell specific Slc7a11 gene knockout and multiple PCR identification technologies, the initial response SPEM state after gastric mucosa damage is successfully simulated, and the kit is used for researching atrophic gastritis pathological mechanisms and evaluating drug intervention.

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 relates to a hepatocellular carcinoma gene knockout target library based on multiple omics and a screening method of the hepatocellular carcinoma gene knockout target library, and the gene knockout target library for precise treatment of hepatocellular carcinoma is finally obtained through data collection and integration, data screening and target verification in sequence. According to the invention, through multi-omics data integration and bioinformatics analysis, key driving genes of hepatocellular carcinoma are systematically screened, and the important effects of the genes in occurrence, development, metastasis, drug resistance and immune escape of hepatocellular carcinoma are disclosed; the genes not only deepen the understanding of the hepatocellular carcinoma molecular mechanism, but also provide important theoretical basis and potential intervention targets for the development of targeted therapy and personalized therapy strategies.

The invention discloses an application of a multidrug-resistant efflux transporter gene OsMRET1 in regulation and control of chalkiness traits of rice. A nucleotide sequence of a coding region of the gene is shown as SEQ ID No.1, and a coded amino acid sequence is shown as SEQ ID No.2. The invention further discloses an application of the multidrug-resistant efflux transporter gene OsMRET1 in regulation and control of chalkiness traits of rice. It is found for the first time that the gene OsMRET1 can regulate and control formation of chalkiness, an OsMRET1 gene knockout mutant is created through a gene editing technology, an OsMRET1 gene overexpression strain is obtained through a transgenic technology, it is verified that the OsMRET1 gene is a new gene for regulating and controlling rice chalkiness, and overexpression of the gene can reduce rice chalkiness and improve rice chalkiness. And a new gene resource and a technical route are provided for rice quality improvement.

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 relates to the technical field of biology, and discloses sgRNA for knocking out a TNIP2 gene, the nucleotide sequence is shown as SEQ ID NO: 1, blocking of the sgRNA to TNIP2 expression is verified through sanger sequencing and Western Blotting experiments, a TNIP2 knockout cell line and a control cell have no obvious morphological and growth difference, and the sgRNA can be used for knockout of the TNIP2 gene. The sgRNA can inhibit the replication ability of the H1 subtype influenza virus by regulating and controlling the expression of a host gene TNIP2, in addition, the invention further discloses application of the sgRNA, a cell strain with the TNIP2 gene knocked out and a construction method of the cell strain.

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 the field of genetic engineering breeding and molecular breeding, and particularly discloses application of ZmMIR319A in regulation and control of heat resistance of corn. According to the invention, corn ZmMIR319A is knocked out by using a gene editing technology, three different types of gene knockout mutants are created, and a ZmMIR319A overexpression material is created at the same time. At the normal growth temperature (25-28 DEG C) of corn, the ZmMIR319A knockout mutant and the overexpression material do not show temperature sensitivity to the wild type, but at the high temperature (45 DEG C), compared with the wild type, the ZmMIR319A mutant is more sensitive to high temperature stress, and the sensitivity of the ZmMIR319A overexpression material to the high temperature stress is reduced, which indicates that the ZmMIR319A positively regulates and controls the high temperature stress resistance of the corn. The invention provides reference for studying the high-temperature-resistant stress molecular mechanism of the corn, and provides potential gene resources for cultivating excellent new germplasm of the high-temperature-resistant corn.

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 relates to the field of bioengineering, in particular to a bilateral cleft lip mouse model based on Pitx2-Cre conditional knockout of Bmp4 as well as a construction method and application of the bilateral cleft lip mouse model. The invention aims to construct a stable bilateral cleft lip mouse model. Experiments find that the Bmp4 gene is expressed in the epithelium of the frontal nose process and the maxillary process at the early lip stage in E9.25. The spatio-temporal expression mode of the Pitx2 gene in the early primordium epithelium is highly consistent with the spatio-temporal expression mode of the Bmp4 gene. Based on this, the inventor constructs a Bmp4 gene knockout model driven by an endogenous Pitx2 gene promoter, and finds that the mouse model shows a 100% bilateral cleft lip phenotype. Compared with the reported Netin-Cre, the Nestin-Cre has the advantages that the Nestin The bilateral cleft lip model constructed by the method is more stable, can better represent genetic diseases of human cleft lip, and has important research significance.

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 belongs to the field of fish breeding, and discloses a construction method and primers of a fast-growing red crucian carp homozygous strain, and the construction method comprises the following steps: preparing gRNA of a red crucian carp acvr2b gene, microinjecting a mixture of the gRNA and Cas9 protein into a one-cell stage fertilized egg of the red crucian carp, and screening to obtain an F0-generation gene knockout mutant; carrying out selfing on the F0-generation gene knockout mutant to obtain an F1-generation mutant, and screening to obtain a homozygous mutant; and selfing and propagating the homozygous mutant to obtain the product. The acvr2b homozygous knockout strain is successfully established in the red crucian carp through a CRISPR / Cas9 gene editing technology for the first time, the body weight of the obtained acvr2b mutant red crucian carp at the age of 12 months is increased by 30.68% compared with that of a wild type, the muscle protein content is increased by 13.81%, the intestinal digestion and absorption capacity is obviously improved, high-quality germplasm resources are provided for fish genetic breeding, and the acvr2b mutant red crucian carp has a good application prospect.

The application relates to the field of genetic engineering breeding and molecular breeding, and particularly discloses ZmMIR319A Application of a gene in regulating a corn leaf angle. ZmMIR319A The gene nucleotide sequence is shown in SEQ ID NO. 1. The application uses a CRISPR / Cas9 gene editing technology to carry out site-specific knockout of a gene in corn ZmMIR319A, Three different types of gene knockout mutants are created, and ZmMIR319A Overexpression materials are created. It is found that, compared with a control, the leaf angle of a leaf on an ear of the knockout mutant is smaller, and the leaf angle of a leaf on an ear of the overexpression plant is increased, which indicates that ZmMIR319A ZmMIR319A The gene can positively regulate a corn leaf angle. The application lays a theoretical foundation for researching a molecular mechanism of a corn leaf angle, and simultaneously provides new molecular markers and gene resources for molecular design breeding and cultivating new excellent corn germplasm with a small leaf angle and a dense planting characteristic.

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.

Disclosed herein are an engineered immune cell with CIITA gene knock-out and use thereof. According to the present invention, an sgRNA specifically targeting CIITA gene is designed and synthesized, which can accurately target CIITA gene to achieve gene knock-out with high knockout efficiency. The provided sgRNA can be used for preparing a universal CAR-T cell.

The invention relates to a method for inducing mouse trophoblast stem cells in vitro. According to the method, mouse embryonic stem cells are used as starting cells, Sorcs3 gene knockout mouse embryonic stem cells are obtained through a gene editing technology, and in-vitro induced trophoblast stem cells (iTSCs) are efficiently and rapidly obtained by means of a flow cytometry sorting technology and in combination with specific antibody sorting. According to the invention, the Sorcs3 gene is knocked out from the embryonic stem cell, so that the capability of the embryonic stem cell to differentiate to the trophoblast lineage can be obviously enhanced; an important research platform is provided for deeply researching a lineage intertransformation mechanism of cell masses (ICM) and trophoblast (TE) in the blastocysts, and the method has important significance for researching lineage limitation of early embryonic development of mammals and the like.

The invention relates to an application of an FOS gene in pig breeding for increasing pig spine number and rib number. The pig capable of increasing the number of spines and the number of ribs is an FOS gene knockout gene editing pig, the number of the FOS gene in an enmbl genome database is ENSSSCG00000002383, the preparation method of the gene editing pig comprises the steps that FOS gene knockout pig fibroblasts are prepared through a CRISPR-Cas9 technology, then the FOS gene knockout pig is produced through a somatic cell cloning technology, and the FOS gene knockout pig is obtained. And finally, selecting pigs with increased spine number and rib number from the FOS gene knockout pigs through phenotype identification. According to the method, the individual proportion of cloned pig fetuses with increased thoracolumbar vertebra number obtained by knocking out the FOS gene is 60%, it is proved that the FOS gene is a key gene influencing the thoracolumbar vertebra number on pig genetic breeding, and a gene edited pig with increased spine number and rib number can be obtained by knocking out the FOS gene, especially knocking out a second exon of the FOS gene.