905 results about "Genome editing" patented technology

The invention discloses a toxoplasma gondii attenuated vaccine strain RH delta rop67 as well as a construction method and application thereof, and belongs to the technical field of parasitic disease prevention and control and biological product preparation. The attenuated vaccine strain is constructed by performing targeted knockout on the ROP67 gene in a toxoplasma gondii strain RH delta ku80 through a CRISPR / Cas9 mediated gene editing technology. Compared with a wild type strain, the attenuated vaccine strain shows remarkable attenuation characteristic and good immunogenicity. A test result shows that the attenuated vaccine strain can induce a host to generate specific immune response mainly based on cellular immunity, maintains a protection effect on toxoplasma gondii infection in a relatively long immune period, and has a protection effect on tachyzoite infection and a chronic infection stage of toxoplasma gondii strains with different virulence; the survival ability of a host to tachyzoite infection can be improved, and the formation level of cysts in tissues is reduced. The invention provides a technical scheme with long-term immune potential for research and development of toxoplasma gondii attenuated vaccines.

The invention discloses a rice salt stress resistant gene OsHAK11 coding protein and application thereof, and belongs to the field of plant genetic engineering. According to the method, a rice OsHAK11 gene (the nucleotide sequence is shown as SEQ ID NO.1) is knocked out through a CRISPR / Cas9 gene editing technology, and a mutant with significantly enhanced salt tolerance is obtained. The gene editing vector pEGCas9Pubi-B-OsHAK11 contains sgRNA of a region as shown in a target SEQ ID NO.4, a rice receptor material is transformed through agrobacterium tumefaciens mediation, and a plant with the OsHAK11 gene subjected to frame shift mutation is obtained through screening. A salt stress experiment shows that the survival rate of the mutant oshak11 is obviously higher than that of a wild type. The invention provides a new gene resource and an efficient technical means for salt-tolerant breeding of rice.

The invention discloses application of an upstream non-coding region uORF of an ABA2 gene in regulation and control of crop agronomic traits, in particular to application in improvement of pre-harvest sprouting resistance of rice and cultivation of pre-harvest sprouting resistant rice strains, and belongs to the technical field of gene engineering. The invention finds that uORF in the upstream non-coding region of the crop ABA2 gene can inhibit the translation level of downstream ABA2 protein on the basis of not influencing the transcriptional level, so that the uORF can participate in regulation and control of crop agronomic traits in which ABA participates in regulation and control, such as improvement of pre-harvest sprouting resistance of rice. Therefore, the upstream non-coding region uORF of the crop ABA2 gene can be modified through a hybridization or gene editing technology, so that the purpose of regulating and controlling crop agronomic traits in which ABA participates in regulation and control is achieved, for example, site-directed mutagenesis is performed on an initiation codon of the upstream non-coding region uORF of the rice ABA2 gene, and the pre-harvest sprouting resistance of rice is improved.

The invention provides a method for constructing a rice epigenetic recombinant inbred line, and belongs to the technical field of molecular biology and genetic breeding. According to the method for constructing the rice epigenetic recombination inbred line, a fertile rice DDM1 gene function weakening mutant is obtained through a CRISPR / Cas9 gene editing technology and hybridized with wild type rice with the same background to obtain F1, then the F1 and the wild type rice are subjected to backcrossing to obtain BC1F1, single-plant offspring selfing with the DDM1 genotype being the wild type is selected, selfing is conducted for multiple generations through a single-grain transmission method, and the rice epigenetic recombination inbred line is obtained. According to the present invention, the strain population with stable inheritance is constructed, such that the rice epigenetic recombinant inbred line is obtained, and has characteristics of rich phenotype, genetic and epigenetic variation so as to provide important population resources for batch identification and the like of important agronomic shape functional sites of rice.

The invention relates to the technical field of gene editing, and particularly provides a method for creating herbicide-resistant rice through gene editing of an OsEPSPS gene promoter region, and the method is characterized in that WT / 29bp single allelic deletion mutation is introduced into the OsEPSPS gene promoter region, i.e., A in an OsEPSPS gene initiation codon ATG is 0 site, A in the OsEPSPS gene initiation codon ATG is 0 site, A in the OsEPSPS gene initiation codon ATG is 0 site, and A in the OsEPSPS gene initiation codon ATG is 0 site; 29bp is deleted from the 112nd site to the 1150th site at the upstream of the initiation codon ATG on one allele, and the other allele is kept unchanged. By introducing the single allelic deletion mutation, the rice can obtain resistance to EPSPS inhibitor herbicides, the resistance is identified in T0-generation plants carrying the mutation and can be stably inherited to offspring plants, and the application value is high for guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

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 belongs to the technical field of plant genetic engineering, and particularly relates to application of a wheat transcription factor TaWRKY22 and a coding gene thereof in regulation and control of plant stripe rust resistance. The function of the wheat transcription factor TaWRKY22 is researched through molecular biology and genetics methods, it is found that knockout of the TaWRKY22 gene significantly improves the resistance of wheat to stripe rust, overexpression of the TaWRKY22 gene reduces the resistance of wheat to stripe rust, and then it is indicated that the wheat transcription factor TaWRKY22 plays a negative regulation role in wheat stripe rust resistance. Therefore, the TaWRKY22 gene is subjected to targeted modification through a gene editing technology, a new genetic material and a molecular breeding strategy can be provided for wheat stripe rust resistance breeding, and an important material and technical support are provided for breeding of wheat disease-resistant varieties.

The invention discloses application of a gene CsPOD7 in cucumber haploid breeding, and belongs to the technical field of molecular breeding. According to the invention, a parthenogenesis haploid induction gene CsPOD7 is cloned from cucumber, the CsPOD7 is knocked out through a CRISPR-Cas9 gene editing technology, and a cspod7 mutant is used as a male parent to be hybridized with other cucumber materials as a female parent, so that parthenogenesis haploid can be generated. The invention is the first application of the gene CsPOD7 in developing a parthenogenesis haploid induction line in dicotyledon, realizes the application of the haploid induction line based on the gene CsPOD7 in cucumber genetic breeding, accelerates the cucumber breeding process, and has huge potential application value and market prospect.

The invention discloses a male sterility gene ZmUCE4 and application thereof in creating a male sterility line of corn, and belongs to the field of plant biotechnology breeding. According to the invention, site-directed mutagenesis of the ZmUCE4 gene in wild type corn is carried out through a CRISPR / Cas9 gene editing technology, pollen development abnormality and abortion can be caused, complete male sterility is caused, and it is proved that the ZmUCE4 gene has a regulation and control function on corn male reproductive development. Through offspring screening, a sterile line without transgenic components can be obtained, a stable maize male sterile line is created, and the method has important significance on maize male fertility control and hybrid seed production. The invention also designs a functional molecular marker aiming at the obtained uce4 male sterility mutant gene, and has important application value in maize male sterility line cultivation, sterile hybrid seed production and molecular marker-assisted selection.

The invention belongs to the technical field of gene engineering, and provides rice purple acid phosphatase OsPAP16 and application thereof, and the application is that a rice purple acid phosphatase OsPAP16 mutant is applied to rice cultivation. Two OsPAP16 mutants are prepared by adopting a gene editing technology, then the mutants are subjected to overexpression induction, and the result shows that the purple acid phosphatase OsPAP16 is positioned on a cell membrane, the OsPAP16 is subjected to phosphorus deficiency induced expression on leaves and roots, the phosphorus deficiency induced expression at the roots is quicker and higher, the overexpression of the OsPAP16 can improve the activity of the acid phosphatase on the root surface, and the yield of the acid phosphatase on the root surface is increased. Phosphorus deficiency stress of the rice can be relieved by degrading organic phosphorus in the environment, the phosphorus content of the rice is increased, and then growth and development of the rice are promoted.

The invention discloses a barley HvNAC92 gene and application of the barley HvNAC92 gene in improvement of plant salt tolerance, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the HvNAC92 gene is as shown in SEQ ID NO. 1. The HvNAC92 gene is knocked out through a CRISPR (clustered regularly interspaced short palindromic repeats) gene editing technology, and the barley mutant with function deletion is successfully obtained. Researches show that the salt tolerance of barley is remarkably enhanced by knocking out the gene, and the growth vigor and biomass of the mutant under the salt stress condition are remarkably superior to those of wild barley. An ion content determination result shows that knockout of the HvNAC92 gene can reduce the transfer of Na ions from a root system to an overground part and increase the absorption of K ions, so that the salt tolerance of plants is improved. Besides, homologous genes of the HvNAC92 gene can be knocked out from crops such as rice, corn, wheat and the like through a gene editing technology, so that the salt tolerance of the plants is improved.

The invention relates to the field of nucleic acid editing, in particular to the technical field of regularly clustered interval short palindromic repeat (CRISPR). In particular, the invention relates to a mutant of a Cas effector protein, a fusion protein comprising the mutant, and nucleic acid molecules encoding the same. The invention also relates to complexes and compositions for nucleic acid editing (e.g., gene or genome editing). The invention also relates to methods for nucleic acid editing (e.g., gene or genome editing) using the proteins or fusion proteins comprising the invention. Compared with a wild type Cas protein, the Cas protein mutant disclosed by the invention has better activity, such as higher cleavage activity, stronger target site recognition capability and higher target sequence editing activity.

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 a transcription factor ZmbZIP27 in regulation and control of corn plant height and ear height, and belongs to the technical field of plant genetic engineering and corn molecular breeding. The invention reveals that the bZIP transcription factor family member ZmbZIP27 has an important biological function of regulating and controlling the corn plant height and lodging resistance for the first time. A ZmbZIP27 knockout vector is constructed through a CRISPR / Cas9 gene editing technology, and an obtained homozygous mutant strain shows remarkable plant type improvement characteristics compared with a wild type: the plant height is reduced by 8-12%, and the ear height is reduced by 15-22%. And a new gene target and a breeding strategy are provided for creating a new high-yield lodging-resistant corn variety suitable for mechanical harvesting.

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 present disclosure provides compositions and methods for altering auxin accumulation in corn or maize plants. Methods and compositions are also provided for altering the expression of genes related to auxin efflux through editing or mutagenesis of a brachytic2 (br2) gene to introduce a premature stop codon or a deletion into the gene such that a truncated Br2 protein encoded by the mutant allele of the br2 gene, which may be a dominant or semi-dominant allele, has at least part of a transmembrane domain without a nucleotide binding domain or motif. Modified plant, plant parts and cells having such a mutant allele with reduced or altered expression or activity of a br2 gene product can have improved characteristics, such as reduced plant height and increased lodging resistance, but without off-types in the plant.

The invention provides an application of a phosphatidylinositol monophosphate 5-kinase gene BnPIP5K9 of rape in regulating and controlling the oil content of the rape. The genetic basis of the oil content of rape seeds is analyzed on the basis of multiple omics, and it is found that the expression quantity of the phosphatidylinositol monophosphate 5-kinase gene BnPIP5K9 in rape is significantly positively correlated with the oil content; the gene BnPIP5K9 is subjected to gene overexpression and knockout to create a mutant for functional verification by further utilizing a tobacco mosaic virus double-35S promoter and a CRISPR / Cas9 gene editing technology, the result shows that the oil content of rape seeds can be remarkably increased by 2.16-2.77% by overexpression of the gene, in addition, rape germplasm resources with gene BnPIP5K9 function deficiency can be obtained through the CRISPR / Cas9 technology, and the rape seed quality is improved. A new theory and a new gene resource are provided for high-oil breeding and oil improvement of the rape, and the method has extremely high application value and potential.

The invention discloses an application of a tobacco cytochrome protein P450 gene, and particularly relates to an application of the tobacco cytochrome protein P450 gene in regulation and control of tobacco dwarfing, and the tobacco cytochrome protein P450 gene is an NtCYP71AU50 gene and / or an NtCYP78A7 gene; the nucleotide sequence of the NtCYP71AU50 gene is as shown in SEQ ID NO. 1, and the nucleotide sequence of the NtCYP78A7 gene is as shown in SEQ ID NO. 3. According to the invention, an editing vector for knocking out NtCYP71AU50 and NtCYP78A7 genes is constructed by virtue of a CRISPR / Cas9 mediated gene editing technology, NtCYP71AU50 and NtCYP78A7 gene editing plants are obtained after genetic transformation, and the plant height, stem girth, waist leaf length, waist leaf width and leaf area of the two gene editing plants are lower than those in a control group. By regulating and controlling the growth and development process of the tobacco plants, the tobacco plants are promoted to be dwarfed, and the problem of shortage of tobacco dwarfing resources is solved, so that application of the tobacco plants in dwarfing close planting cultivation is promoted, the lodging resistance of the tobacco is enhanced, and possibility is provided for increasing the yield per unit area.

The invention relates to an application of a genetic transformation method of populus sibirica and a CRISPR / Cas9 gene editing method in plant breeding, and belongs to the technical field of plant molecular breeding. The method comprises the following steps: (1) soaking an explant in an infection bacterium solution; (2) spreading on a CM1 culture medium; (3) transferring the explant to a CM2 culture medium for 4-5 weeks; (4) transferring the calluses to a CM3 culture medium until adventitious buds are obtained; (5) transferring the callus with the adventitious buds to a CM4 culture medium until cluster buds are obtained; and (6) cutting off clustered seedlings, and inserting the clustered seedlings on a CM5 culture medium. According to the application of the genetic transformation and CRISPR / Cas9 gene editing method of the poplar in plant breeding, a target gene is transferred into a poplar body to obtain a positive plant, meanwhile, a poplar phytoene dehydrogenase (PDS) gene is taken as an example, a poplar mutant of which the PDS gene is knocked out is successfully obtained by applying a CRISPR / Cas9 gene editing technology, and the method is suitable for plant breeding. And a technical support is provided for application of a CRISPR / Cas9 gene editing technology in molecular breeding of the populus sibirica.

The invention relates to a male gamete killer gene S19g-A1 of a rice interspecific hybrid sterile S19 seat and application of the male gamete killer gene S19g-A1. The invention provides a rice hybrid sterility related protein S19g-A1. The amino acid sequence of the rice hybrid sterility related protein S19g-A1 is as shown in SEQ ID NO. 2. According to the invention, a key gene S19g-A1 of a rice interspecific hybrid sterile S19 site is researched and identified, function knockout is carried out on African rice S19g-A1, and pollen fertility of a mutant hybrid F1 obtained by carrying out distant hybridization on an obtained African rice s19g-a1 mutant returns to normal. Based on a gene editing technology, the invention develops a method for rapidly creating a hybrid affinity line capable of overcoming sterility of African rice hybrid, effectively eliminates S19 locus mediated interspecific hybrid reproductive disorder, breaks through the bottleneck of rice interspecific hybrid advantage utilization, provides target gene resources for distant hybrid advantage utilization and crop yield improvement, and has a wide application prospect. Good application prospects are realized.

The invention belongs to the technical field of gene editing, and particularly relates to a construction method and application of a needle mushroom CRISPR / Cas9 gene editing vector. The CRISPR / Cas9 gene editing vector for flammulina velutipes is characterized in that the vector takes pFgnpt as a skeleton vector, and further comprises a Cas9 expression cassette and an sgRNA expression cassette; the nucleotide sequence of the Cas9 expression cassette is as shown in SEQ ID NO. 10. The nucleotide sequence of the sgRNA expression cassette is as shown in SEQ ID NO. 11. According to the present invention, with the agrobacterium-mediated transformation technology, the stable expression of Cas9 and sgRNA in the flammulina velutipes body is achieved, the efficient and stable CRISPR / Cas9 gene editing system is established, and the technical support is provided for the flammulina velutipes variety improvement, the functional gene research and the industrial upgrading.

The invention relates to the technical field of rice gene engineering, in particular to application of a rice gene OsbZIP21 in regulation and control of cold resistance. The invention discloses an application of a rice gene OsbZIP21 in regulation and control of cold resistance. The nucleotide sequence of the OsbZIP21 gene is as shown in a sequence table SEQ ID NO: 1, and the coded amino acid sequence is as shown in a sequence table SEQ ID NO: 2. According to the invention, the rice OsbZIP21 gene is edited by adopting a CRISPR / Cas9 gene editing technology, the expression of the encoding protein of the rice OsbZIP21 gene is silenced, and two forms of cold-resistant rice mutants are obtained. Experimental results show that under the condition of cold stress, the survival rate of the two obtained rice mutants is obviously higher than that of wild rice mutants, and it is indicated that the OsbZIP21 gene plays an important role in regulation of the cold tolerance of rice. The invention provides new resources and technical means for cold-resistant breeding of rice and other plants.

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 discloses a cell lineage tracking system based on synNotch and CRISPR / Cas9 bar code technology and application thereof, the cell lineage tracking system comprises a system mGFP ligand vector for constructing Sender ligand cells and a system for constructing Receiver recipient cells, and the system comprises a synNotch receptor-rtTA fusion vector, a TetO-Cas9 expression vector, a gRNA expression vector and a Target vector library containing a Barcode sequence. A synNotch system and a CRISPR / Cas9 gene editing technology are combined with a bar code strategy, a unique bar code is generated, the contact sequence between cells is recorded, the method is compatible with a single-cell sequencing technology, transcription information of the cells is provided, long-term tracking of the contact history between the cells is achieved, the method can be used for tracking interaction and functions between the cells in a tumor microenvironment, and the application prospect is wide. Through long-term tracking and functional analysis of interaction between macrophages, especially macrophages and tumor cells, the accuracy of interaction analysis is remarkably improved, and a more accurate target spot is provided for targeted therapy.

The invention relates to the field of nucleic acid editing, in particular to the technical field of regularly clustered interval short palindromic repeat (CRISPR). In particular, the invention relates to a mutant of a Cas effector protein, a fusion protein comprising the mutant, and nucleic acid molecules encoding the same. The invention also relates to complexes and compositions for nucleic acid editing (e.g., gene or genome editing). The invention also relates to methods for nucleic acid editing (e.g., gene or genome editing) using the proteins or fusion proteins comprising the invention. Compared with a wild type Cas protein, the Cas protein mutant disclosed by the invention has better activity, such as higher cleavage activity, stronger target site recognition capability and higher target sequence editing activity.

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 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 invention relates to a haploid induction method based on a brassica napus phospholipase gene BnaPLA2-a and application of the haploid induction method, the BnaPLA2-a gene in brassica napus is knocked out through a gene editing technology, a mutant material capable of inducing haploid generation is obtained, and the application blank of a phospholipase pathway in dicotyledon haploid induction is filled. Compared with a traditional microspore culture technology, the invention provides a brand-new haploid induction path which is derived from the rape and is used for the rape. The invention provides a brand new technical tool and germplasm resources for genetic breeding of brassica napus. Meanwhile, the invention discloses a gene, a mutant, a creation method and application in breeding.

Disclosed herein are methods for rapid transformation, genome editing, and regeneration of dicot plants. The methods disclosed herein involve transforming plant cells and explants with recombinant DNA to achieve genome edits. The methods enhance transformation and regeneration efficiency and reduce the time to produce transgenic plants.

The invention provides application of an MsMYBS3 gene or a biological material thereof in regulation and control of plant cold resistance, and belongs to the technical field of plant genetic engineering. The amino acid sequence coded by the MsMYBS3 gene disclosed by the invention is as shown in SEQ ID NO. 4. The invention provides an application of a medicago sativa MYB transcription factor gene MsMYBS3 in the aspect of regulating and controlling the cold resistance of medicago sativa. A gene editing technology is adopted to directionally change the development and the cold resistance of the medicago sativa. The cold resistance of an MsMYBS3 gene editing strain constructed by the invention is reduced, and the MsMYBS3 gene has the effect of regulating and controlling the cold resistance of alfalfa, so that a theoretical basis and a candidate gene are provided for cultivating new varieties of high-resistance leguminous forage, and the application of alfalfa in production is promoted.