585 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 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 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 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 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 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.

The invention discloses a high-efficiency wide-targeting-range C-to-G double-enzyme synergistic base editor and application thereof, and belongs to the technical field of gene editing. The editor is a fusion protein comprising a Cas protein having reduced or lost endonuclease activity, a cytosine deaminase (CDA), and a cytosine DNA glycosylase (CDG). In order to solve the problems that an existing C-to-G editor is low in efficiency and limited in targeting range, the C-to-G editor synergistically and efficiently generates a base removal (AP) site on a target DNA through the dual effects of CDA and CDG, so that C-to-G base transversion is promoted. According to the editor, the C-to-G editing efficiency is remarkably improved, an editing window can be effectively expanded or moved, the targeting flexibility is greatly enhanced, meanwhile, high genome specificity is kept, and the editor shows strong application potential in various organisms such as yeast and plants.

This invention belongs to the field of molecular genetics and discloses a method and application for controlling fish fertility using in vivo gene editing technology. The applicant, for the first time, cloned a carp-specific U6 promoter and used this promoter to construct the U6 gene. MOVIE gRNA transgenic vectors and water The Cas9 vector allows for the establishment of an in vivo editing system. This strategy enables the heritability of sterility, a reproductive control trait, through hybridization of fertile parents to produce sterile offspring. The operation is highly efficient, simple, and environmentally friendly. Furthermore, since high-copy-rate short, scattered repetitive sequences exist in all fish species, this strategy has broad applicability across species.

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.

This invention belongs to the fields of locust biotechnology and gene editing technology, specifically relating to a method for preparing locusts. Crz Methods using gene mutants and their application in locust control. This invention discovers that inhibiting gene mutants in locusts... Crz Gene expression can be used to control locusts. Crz The gene sequence is shown in SEQ ID NO:3. This invention successfully established a homozygous Crz mutant strain of the migratory locust using gene editing technology. It was found that the albino migratory locusts obtained after Crz deletion were more easily preyed upon by natural enemies than the wild-type locusts, exhibited significantly lower levels of the aggregation pheromone 4VA, and showed weaker adaptability to low temperatures compared to the wild type. This provides a new strategy for the biological control of migratory locusts and can also serve as a reference for the biological control of other pests.

The application discloses OsHAK13 Application of a gene in regulating drought tolerance of rice relates to the technical field of genetic engineering. OsHAK13 The gene encodes an amino acid sequence as shown in SEQ ID NO. 2. OsHAK13 By knocking out the gene through a gene editing technology, a functional loss mutant is obtained, which shows significant enhancement of drought tolerance at a seedling stage. The gene and the mutant can be applied to genetic improvement of crop drought resistance, and provide important gene resources and breeding materials for cultivation of new water-saving and drought-resistant rice varieties, and have important theoretical significance and application value.

The invention belongs to the technical field of crop breeding by a gene editing technology, and particularly relates to a method for creating a novel tomato cytoplasmic male sterility germplasm through gene editing and an application of the novel tomato cytoplasmic male sterility germplasm. In order to solve the problems that existing cytoplasmic male sterile materials in tomatoes are insufficient, production of tomato hybrid seeds is restricted and the like, gene editing is performed on MSH1 through a CRISPR / Cas9 technology, cytoplasmic male sterile single plants are identified in mutant selfing separation progenies and hybridized with wild types twice, and the tomato hybrid seeds are obtained. Finally, a novel non-transgenic tomato cytoplasmic male sterility material with the MSH1 gene recovered to a wild type is obtained, and meanwhile, the horticultural character of the original fertile material is kept unchanged.

This invention discloses a genetic transformation and gene editing method applicable to multiple sesquiterpene species, belonging to the field of plant genetic transformation and gene editing technology. The method uses the cotyledonary nodes of sterile seedlings of the sesquiterpene genus as explants, and introduces an optimized pScEF1α-Cas9 gene editing transformation vector using Agrobacterium infection. Regenerated plants are obtained through recovery culture, selection culture, and rooting culture. The vector contains four gene expression units: pScEF1α:Cas9, pGmU6:sgRNA, pGmUBI:DsRed2, and pCaMV35S:HygR. In vivo screening is further performed using the DsRed2 fluorescent reporter gene, and the gene-edited plants are identified by molecular detection. This method is applicable to common sesquiterpene, stem-nodled sesquiterpene, and spiny sesquiterpene, and has advantages such as strong species versatility, high transformation efficiency, short cycle, visualized screening, and high positive rate, providing efficient technical support for gene function research and genetic improvement of sesquiterpene species.

The application belongs to the technical field of biological gene engineering, and particularly relates to a barley HvCslf6 promoter element and application thereof in regulating the content of barley grain beta-glucan, wherein the barley HvCslf6 promoter element is located at 295bp-300bp upstream of a transcription start site of a barley HvCslf6 gene; the nucleotide sequence of the HvCslf6 promoter element is CCGTTG, and the HvCslf6 promoter element belongs to a CCAAT-box element; the gene number of the HvCslf6 gene in BARLEX Morex v3 Gene Models is HORVU.MOREX.r3.7HG0698110. The application finds a key element which can affect the content of beta-glucan in barley grains in the upstream promoter region of a main effect gene HvCslf6 of barley beta-glucan synthesis by using a gene editing technology, and thus provides a method capable of preparing transgenic plants with changed content of beta-glucan in barley grains, and provides a new means and tool for detecting and regulating the content of beta-glucan in barley grains.

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.

This invention discloses the application of knocking down or knocking out the OsABA8ox2 protein or its encoding gene in improving the panicle germination resistance of rice seeds, belonging to the fields of biotechnology and plant genetic engineering technology. OsABA8ox2 The amino acid sequence of the protein is shown in SEQ ID NO.2, and the nucleotide sequence of the gene is shown in SEQ ID NO.1. This invention utilizes CRISPR / Cas9 technology to directionally edit the target site sequence in this gene, as shown in SEQ ID NO.3, successfully obtaining rice nucleotide sequences as shown in SEQ ID NO.8, SEQ ID NO.9, and SEQ ID NO.10. OsABA8ox2 The mutant, to a certain extent, enhances the dormancy of rice seeds, effectively reducing the germination rate of rice panicles. The gene resources and gene editing technology provided by this invention offer an important technical approach for rapidly improving or creating rice varieties with significantly enhanced panicle germination resistance in agricultural production, demonstrating significant application value in production practice.

Disclosed is a method for editing mitochondrial DNA (mtDNA) within a cell, which include introducing into the cell (a) a DNA cleaving enzyme targeted to the mtDNA sequence to be deleted; (b) a first DNA binding component targeted to a sequence adjacent to the 5′ end of a mtDNA sequence to be deleted; and (c) a second DNA binding component targeted to a sequence adjacent to the 3′ end of the mtDNA sequence to be deleted, where the DNA cleaving enzyme generates a double stranded break (DSB) within the mtDNA sequence to be deleted or generates a single strand nick on the light strand of the mtDNA sequence to be deleted, and wherein the mtDNA sequence between the target sequence for the first DNA binding component and the target sequence for the second DNA binding component is deleted.

The application belongs to the technical field of genetic engineering, and particularly relates to application of a GadW mutant in human milk oligosaccharide production. The amino acid sequence of the GadW mutant is shown as SEQ ID NO. 3. By using a gene editing technology, a genetic engineering chassis strain carrying a GadW mutant coding gene is obtained, and then the above chassis strain is applied in a human milk oligosaccharide such as 3'-SL, 6'-SL, LNT II, LNnT and LNT, etc. production strain construction process, and the yield of human milk oligosaccharide is further improved. Among them, the yield of 3'-SL is increased by 8.5%, the yield of LNT II is increased by 26.7%, the yield of LNnT is increased by 50%, the yield of LNT is increased by 42.9%, and the yield of 6'-SL is increased by 35.3%.

The invention relates to the technical field of gene editing, and discloses an sgRNA activity prediction method and device based on deep features and automatic machine learning. The method comprises the following steps: acquiring sgRNA training data with editing efficiency labels and discretizing the sgRNA training data into classification labels; after the sgRNA and the PAM sequence are spliced, inputting the spliced sgRNA and PAM sequence into a pre-training depth sequence model to extract high-dimensional features; obtaining a low-dimensional feature vector through nonlinear dimensionality reduction; an automatic machine learning framework is combined with cross validation, and a prediction model is automatically trained and optimized based on low-dimensional features; and finally, carrying out activity prediction and sorting screening on candidate sgRNA by utilizing the model. According to the method, through fusion of deep semantic extraction, nonlinear structure maintenance and a full-automatic modeling process, the accuracy, robustness and development efficiency of sgRNA activity prediction are effectively improved, and an efficient pilot screening tool is provided for a gene editing experiment.

The present application belongs to the field of plant genetic engineering and biotechnology, and particularly relates to a method for creating a dwarf material of Brassica napus by using CRISPR / Cas9 gene editing technology and application. The method edits the Brassica napus BnaA06G0083400WE gene by using CRISPR / Cas9 gene editing technology to obtain a dwarf material of Brassica napus; the nucleotide sequence of the coding gene mutant of the Brassica napus dwarf material is SEQ ID NO. 1. The present application edits the Brassica napus BnaA06G0083400WE gene by using CRISPR / Cas9 gene editing technology to obtain a dwarf Brassica napus, which provides valuable genetic resources and germplasm resources for Brassica napus breeding; the method has strong characteristics and is easy to obtain, is an effective way to realize the improvement of target traits and cultivate new materials, and can be applied to Camellia sinensis breeding.

The application belongs to the technical field of gene editing, and particularly relates to an optimized MmeFz2 protein and gene editing application thereof. The optimized MmeFz2 protein is obtained by mutating a wild-type MmeFz2 protein shown in SEQ ID NO. 1 as follows: mutating C at the 69th position of the wild-type MmeFz2 protein into K, mutating E at the 305th position into N and mutating E at the 326th position into Q; or mutating E at the 178th position of the wild-type MmeFz2 protein into H, mutating E at the 305th position into S and mutating E at the 418th position into R. The C-terminal of the optimized MmeFz2 protein is further fused with an HMG-D protein. The application further provides an optimized MmeFz2-omega RNA system, and the system has strong gene editing efficiency. The application has important significance and value for promoting the wide application of gene editing technology in functional mechanism research and clinical treatment.

This invention relates to the field of gene editing technology, specifically providing a method for creating herbicide-resistant rice by gene editing the promoter region of the OsEPSPS gene. The method involves introducing a WT / -29bp monoallelic deletion mutation into the OsEPSPS gene promoter region. Specifically, with position A of the OsEPSPS gene start codon ATG set to 0, a 29bp deletion is made in one allele located at positions 1122 to 1150 upstream of the start codon ATG, while the other allele remains unchanged. This invention demonstrates that by introducing this monoallelic deletion mutation, rice can acquire resistance to EPSPS inhibitor herbicides. This resistance was identified in the T0 generation plants carrying this mutation and can be stably inherited by their offspring, possessing significant application value for ensuring agricultural production safety and improving herbicide application efficiency.

This invention pertains to fusion protein mutants comprising a Prime Editing enzyme having a first amino acid sequence and a second amino acid sequence, wherein the first amino acid sequence comprises a SpCas9 H840A nickase mutant protein of SEQ ID NO:152 and the second amino acid sequence comprises a Moloney Murine Leukemia Virus reverse transcriptase protein mutant (MMLV RTase mutant), wherein the fusion protein mutant displays at least the equivalent or greater activity of a reference Prime Editing enzyme in genome editing.

The invention belongs to the technical field of gene editing, and particularly relates to a WER10 enhancer and application thereof. The invention provides application of a WER10 nucleic acid molecule and / or a biological material for regulating and controlling the WER10 nucleic acid molecule, the nucleotide sequence of the WER10 nucleic acid molecule is as shown in SEQ ID NO: 1, and the application comprises any one or more of the following items: 1) application of the sequence as shown in SEQ ID NO: 1 as an enhancer; 2) application in enhancing the expression of a downstream reporter gene LUC induced by an immune induction signal; and 3) application in regulation and control of the broad-spectrum antibacterial property of the plants. The sequence and the characteristics of the pathogenic bacterium induced enhancer WER10 located in the intergene region are identified and verified from arabidopsis thaliana, plant immunity related cis-regulatory element resources are enriched, and a new resource is provided for constructing a pathogen induced disease-resistant expression system and developing crop disease-resistant molecular design and breeding.

CRISPR-Cas systems have been engineered for various purposes, such as genomic DNA cleavage, base editing, epigenome editing, and genomic imaging. Although significant developments have been made, there still remains a need for new and useful CRISPR-Cas systems as powerful precise genome targeting tools. The invention disclosed herein comprises CRISPR-Cas based compositions and methods for high integration efficiency and expression efficiency of transgenes together with high post-transfection cell viability in eukaryotic cells.

The present invention relates to a novel polynucleotide applicable to genome editing techniques with improved editing efficiency of a target nucleotide sequence. The method using the double-stranded nucleotide complex of the present invention enables modification of a target nucleotide sequence solely by introducing the double-stranded nucleotide complex, without introducing an exogenous nuclease or a gene encoding an exogenous nuclease into a cell, and is thus applicable to genome editing techniques with extremely high safety.

Embodiments disclosed herein include novel nucleic acid-guided nucleases, novel guide nucleic acids, and novel targetable nuclease systems, and methods of use. In some embodiments, engineered non-naturally occurring nucleic acid-guided nucleases, can be used with known guide nucleic acids in a targetable nuclease system. In certain embodiments, targetable nuclease systems can be used to edit targeted genomes of humans and other species. In some embodiments, methods include, but are not limited to, recursive genetic engineering and trackable genetic engineering methods.

This invention relates to the field of animal model construction technology, and more particularly to a method for constructing an SMDT1 gene knockout animal model. The method utilizes gene editing technology to enable the animal model to... SMDT1 The sequence shown in SEQ ID No. 1 is missing from the locus. Stable and reliable results can be obtained using the construction method of this invention. SMDT1 Gene knockout animal models do not exhibit recombination repair and can be stably inherited, making them suitable for research. SMDT1 The biological functions of genes and MCU complexes, and their potential applications in... SMDT1 Gene-targeted drug development and guidance for livestock breeding have broad application prospects.