1224 results about "Gene knockout" patented technology

The present invention relates to the fields of genetic engineering and plant genetic breeding, and specifically to the application of wheat TaPND gene. In the present invention, by taking gene TaPND as a target, designing a sgRNA sequence based on CRISPR / Cas9, connecting a DNA fragment containing the encoding sgRNA sequence to a vector carrying CRISPR / Cas, a plant binary expression vector capable of knocking out the TaPND gene in wheat is constructed; by genetically transforming wheat, offspring with TaPND gene knockout are obtained, and wheat mutants with early flowering time are obtained. Experiments have confirmed that transgenic wheat with TaPND gene knockout achieves the advancement of wheat flowering time. The technical scheme of the present invention provides a practical method for realizing rapid wheat breeding using genetic engineering technology, which has important breeding application value and broad market application prospects.

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 a porcine muscle stem cell line capable of being cultured in vitro for a long time and application of the porcine muscle stem cell line, and belongs to the technical field of stem cells and cell culture meat. The porcine muscle stem cell line is prepared by the following steps: performing CDKN2A gene knockout on porcine muscle stem cells by using a CRISPR / Cas9 gene editing technology, performing monoclonal screening and genotype identification to obtain a cell strain with successful CDKN2A knockout, and then performing carrier-free and serum-free suspension culture domestication on the cell strain. And a carrier-free and serum-free porcine muscle stem cell line capable of being subjected to in-vitro long-term suspension culture is obtained. Verification shows that the CDKN2A knockout porcine muscle stem cell line subjected to carrier-free and serum-free suspension culture domestication not only can be subjected to carrier-free and serum-free suspension culture in a shake flask for a long time, but also can maintain a good differentiation characteristic. The porcine muscle stem cell line which can be cultured on a large scale, is derived from edible animals and can be cultured in vitro for a long time is created, and excellent cell resources are provided for commercial production of cultured meat.

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 discloses a cotton GhSNL1 protein as well as a coding gene and application thereof. The cotton GhSNL1 protein coding gene (GhA01G1757; according to the present invention, the CRISPR-Cas9 gene editing material of the GhSNL1 gene is subjected to phenotypic analysis to discover that the GhSNL1 gene knockout material has the phenotype of fiber start slowing, fiber density shortening and mature plant boll regeneration, such that the GhSNL1 gene participates in the cotton fiber development process, and the cotton fiber development efficiency is significantly improved; the method provides a theoretical basis for cotton variety improvement, and can be used for cultivating new cotton varieties with excellent fiber quality and high yield.

The invention belongs to the technical field of molecular biology, and particularly relates to sphingomonas paucimobilis recombinant engineering bacteria as well as a construction method and application thereof. The engineering strain is obtained by knocking out a metabolic bypass gene from sphingomonas paucimobilis, and the metabolic bypass gene is composed of a crtI gene or a phaC gene. The production amount of gellan gum can be increased by the delta phaC knockout strain and reaches 8.9 g / L. In addition, overexpression of energy metabolism related genes pgi or clpB or ycaJ is carried out, and the yield of a strain with the overexpressed pgi gene is increased to the maximum and reaches 9.5 g / L. Therefore, the pgi gene is overexpressed on the basis of the delta phaC knockout strain, the yield of gellan gum is further increased to 10.0 g / L, and the production efficiency is improved.

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 a gene editing vector of agrobacterium, a gene editing method of the gene editing vector and an agrobacterium editing strain. The agrobacterium gene editing vector comprises the following gene elements: a plasmid skeleton, a Pvan promoter, a spCas9 gene, a green fluorescent protein gene GFP, gRNA, and an upstream homologous arm and a downstream homologous arm of a target gene. According to the agrobacterium gene editing vector, a CRISPR / spCas9-GFP gene editing system is adopted, an efficient, simple, convenient and traceless genome editing system is constructed in agrobacterium, and gene knockout and insertion can be achieved.

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.

The invention relates to a recombinant pichia pastoris genetically engineered bacterium for expressing recombinant full-chain-length humanized collagen as well as a preparation method and application of the recombinant pichia pastoris genetically engineered bacterium, and belongs to the technical field of genetic engineering. The recombinant pichia pastoris genetically engineered bacterium disclosed by the invention expresses recombinant full-chain-length humanized collagen, and a ku70 gene of the recombinant pichia pastoris genetically engineered bacterium is knocked out and knocked into a recombinant 4-proline hydroxylase encoding gene. Recombinant full-chain-length humanized collagen is optimized, a 4-proline hydroxylase gene and a collagen gene are co-expressed in pichia pastoris to obtain hydroxylation modified recombinant collagen, and a corresponding recombinant pichia pastoris genetically engineered bacterium is provided, has proteolytic enzyme A defect and hydroxylation modification capability, and can be used for preparing a recombinant collagen product. The stability of the collagen in the fermentation process is improved. The recombinant full-chain-length humanized collagen provided by the invention is 100% homologous with III-type collagen derived from a human body, so that the immune risk is extremely low, and the biological activity is more perfect.

The invention provides a wheat drought-tolerant gene TaTZF1 and application thereof in plant drought tolerance, and belongs to the technical field of gene engineering. The invention provides a wheat drought-enduring gene TaTZF1. The amino acid sequence of a protein encoded by the wheat drought-enduring gene TaTZF1 is as shown in SEQ ID No. 2. The wheat drought-enduring gene TaTZF1 is subjected to induced expression by drought, ABA and salt stress, the wheat drought-enduring gene TaTZF1 is overexpressed and transformed into wheat in an overexpression mode, and it is found that the drought tolerance of a transgenic plant with overexpressed TaTZF1 is remarkably improved compared with that of a wild type plant; after the wheat drought-tolerant gene TaTZF1 in wheat is knocked out, the drought tolerance is obviously reduced. It is proved that the wheat drought-resistant gene TaTZF1 positively regulates the drought resistance of wheat, gene resources are provided for wheat molecular breeding, and the wheat drought-resistant gene TaTZF1 has great significance in improvement of wheat drought-resistant genetic improvement.

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 relates to the technical field of biology, in particular to a recombinant strain for producing nicotinamide mononucleotide and application of the recombinant strain. According to the method, accumulation of the NMN is improved through metabolic flux redirection and gene knockout, a synthesis path of the NMN is successfully established in escherichia coli, and further, the yield of the NMN is effectively improved and reaches 7.0 g / L by optimizing a transport system of NAM-NMN and a PRPP synthesis path. The invention also optimizes the production process of the NMN, realizes the highest yield of the NMN of 28.5 g / L in a fed-batch fermentation system, and obviously improves the yield compared with the traditional batch fermentation.

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 provides a coding protein of a rice blast-resistant super-effect allele ETD1. The protein has a nucleotide sequence as shown in SEQ ID NO.3. The invention also provides a preparation method of the rice blast-resistant super-effect allele ETD1. The coding gene of the protein belongs to a super-efficient allele of OsCNGC13, obtains stronger calcium ion transport capacity, and has a function of remarkably accelerating the influx of cell calcium ions. Therefore, the rice resistance can be improved by normal expression or overexpression of the protein in a gene knockout type plant, so that the rice blast resistance can be positively regulated and controlled.

The invention provides an application of a Brassica napus gene BnSKIP31 in prevention and control of sclerotiniose, which is an application in obtaining a disease-resistant enhanced rape material by creating a gene to edit rape. According to the invention, CRISPR gene knockout rape of BnSKIP31 is constructed, the negative regulation function of the gene on sclerotiniose resistance is disclosed for the first time, and application of the BnSKIP31 gene to acquisition of sclerotiniose rape materials with increased sclerotiniose resistance by creating CRISPR gene knockout rape is provided. The BnSKIP31 gene provided by the invention is a new gene resource suitable for creating and breeding new sclerotiniose-resistant rape materials and new varieties, and the created rape germplasm has lasting disease resistance.

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 provides a method for synthesizing D-psicose through fermentation of escherichia coli by using D-glucose and D-xylose. The method comprises the following steps: by taking escherichia coli as a host cell, over-expressing a galactose permease gene galP, a glucokinase gene glk, a 6-phosphate glucose isomerase gene pgi, a D-psicose-6-phosphate epimerase gene alsE with an SUMO tag and a D-psicose-6-phosphate phosphatase gene a6PP, and constructing a path for synthesizing D-psicose by taking D-glucose and D-xylose as substrates; the synthetic route of D-psicose is optimized by knocking out a D-glucose phosphorylation transport pathway gene ptsG, knocking out D-fructose-6-phosphokinase genes pfkA and pfkB, silencing a glucose-6-phosphate dehydrogenase gene zwf, overexpressing glutamate transhydrogenase genes gdh1 and gdh2, knocking out a UDP-glucose-4-epimerase gene galE and knocking out a transport protein key gene fryA in D-psicose; and finally, the optimal proportion of mixed sugar fermentation is explored, so that efficient synthesis of D-psicose from escherichia coli through mixed sugar fermentation of D-glucose and D-xylose is realized.

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 discloses construction and application of a streptococcus suis serotype 2 cps2C gene knockout mutant strain. The invention relates to a streptococcus suis serotype 2 cps2C gene knockout mutant strain 05ZYH33 delta cps2C. A coding gene, from the 31st site to the 1000th site, of the cps2C gene in the strain 05ZYH33 is replaced by a spectinomycin resistance gene cassette SpcR. The mutant strain 05ZYH33 delta cps2C is applied to preparation of a streptococcus suis type 2 attenuated vaccine and a subunit vaccine. The capsule of the mutant strain 05ZYH33 delta cps2C is obviously reduced; in-vitro experiments show that the macrophage phagocytosis resistance of the mutant strain 05ZYH33 delta cps2C is reduced, and animal toxicity test results show that the toxicity of the mutant strain is obviously reduced. The mutant strain provides an important clue for screening of protective antigens of multivalent subunit vaccines, and can be applied to development of S.suis attenuated vaccines and multivalent subunit vaccines.