49 results about "Plant genomes" patented technology

The invention discloses application of novel reverse transcriptase and related fusion protein of the novel reverse transcriptase in plant-guided editing. Candidate RT protein is obtained through systematic mining from a public database, novel reverse transcriptase cl1 with activity is screened out in combination with a fluorescence report system, and a guided editing system suitable for plants is constructed. The system tests in wheat protoplast, and verifies the editing activity of cl1 to a plurality of endogenous targets. Further combining with protein structure prediction and rational design point mutation optimization, the cl1 mutant with higher editing efficiency is obtained, and the method can be applied to accurate and effective editing of plant genomes and plant breeding and improvement.

The invention belongs to the field of gene engineering, and relates to nuclease generated based on AI and having a gene editing function and application of the nuclease. The technical problem to be solved by the invention is to develop more nuclease suitable for eukaryote gene editing. According to the technical scheme, nuclease AIGCprotein which is generated based on AI and has a gene editing function is obtained through the following steps that a plurality of protein amino acid sequences are generated through a known nuclease three-dimensional structure in a PDB database, the sequences are classified and compared, and a protein skeleton of the nuclease AIGCprotein is obtained through extraction; the known nuclease is Cas12. The invention provides nuclease which is generated on the basis of Protein MPNN AI and has a gene editing function, and the protein does not exist in the nature; the method can be used for constructing a plant genome editing system.

The present invention relates to a method for preparing a MAGEA8-Fc transgenic plant. Through a platform using a tobacco plant, a recombinant protein MAGEA8-Fc, in which a human IgG Fc fragment is fused to a MAGEA8 protein, can be stably expressed. The method of the present invention enables high-level expression efficiency and stable protein production by introducing a target gene into a plant genome using an Agrobacterium binary vector. By directly isolating and purifying the recombinant protein from the transgenic tobacco plant, large-scale production can be easily achieved. Compared to conventional microbial or animal cell-based expression systems, the method provides economic efficiency and scalability, and thus can be usefully applied as a platform for producing fusion proteins.

Methods arc provided for making a plant genome having a desired nucleic acid sequence. One example method includes identifying genomic segments of a plant that include at least one desired nucleic acid sequence, using a trained neural network, which is biology-informed through first pre-training connections between a genetic data layer and at least one intermediate layer and second pre-training connection between the at least one intermediate layer and a phenotypic layer. The method also includes recombining the at least one at least one desired nucleic acid sequence with a second nucleic acid sequence to form a target plant genome.

The invention belongs to the field of gene engineering. Specifically, the invention relates to a method for inserting an exogenous sequence in a genome at a fixed point. Specifically, on the basis of a guided editing system (PE), two adjacent pegRNAs with partially overlapped sequences on a reverse transcription template are used, and efficient and accurate exogenous sequence fixed-point insertion is realized in a genome, especially a plant genome. The system is further coupled with a recombinase system such as Cre / Lox or FLP / FRT and the like, and large-fragment exogenous sequence fixed-point insertion is realized in genomes, especially plant genomes.

The application discloses a kind of plant whole genome RNA-chromatin interaction detection methods, the method includes the interaction state of fixed RNA and chromatin, preparation double link, obtain nuclear, chromatin DNA fragment, RNA / linker / DNA chimera, RNA-DNA double-stranded complex in turn;Tn5 transposase fragmentization cuts double-stranded complex;PCR amplification forms DNA library.The application applies Tn5 transposase to the original RADICL-seq method, successfully to RNA-DNA complex is cut and library, greatly simplifies the process, experimental procedure is reduced by at least half, experimental period is shortened from 4-5 days to 3 days, the fragment size of DNA library produced is between 250bp-400bp, increase the length of final effective RNA / DNA fragment pair, thereby greatly improve the efficiency when aligning to plant genome in subsequent bioinformatics analysis, provide effective means for plant whole genome RNA-chromatin interaction detection.

Disclosed herein are methods of improving agronomic traits in crop plants through editing sequences within the 5′ untranslated region (UTR) of plant genomes.

The invention belongs to the technical field of agricultural biology, and particularly relates to a rice endogenous space-time specific promoter OsSULTR1; the invention discloses 3pro and application thereof. The promoter OsSULTR1 is characterized in that the promoter OsSULTR1 The nucleotide sequence of 3pro is as shown in SEQ ID NO. 1. The invention provides an OsSULTR1; 3pro is a space-time specific expression regulation element from rice, can efficiently and stably drive a target gene to be expressed in a plant callus, and can realize targeted efficient and stable expression in root and stem tissues in a plant vegetative growth period. The promoter can effectively replace a non-plant-source promoter commonly used in existing plant genetic engineering, has an important industrial application value in the field of plant genetic engineering, and not only can improve the targeting and stability of target gene expression in transgenic plants, but also can improve the yield of the transgenic plants. And the potential biological safety risk of the transgenic plant caused after the exogenous DNA is introduced into the plant genome can be obviously reduced.

The invention relates to the technical field of agriculture, in particular to an efficient CBE base editing tool based on Cas12i3 and application of the efficient CBE base editing tool. According to the Cas12i3-based efficient CBE base editing tool provided by the invention, the base editing tool comprises a protein expression cassette; the nucleotide sequence of the protein expression cassette is as shown in SEQ ID NO. 1. The tool can realize efficient, accurate and low-off-target C-to-T single base replacement in a plant genome, the editing efficiency and the target flexibility are remarkably improved, and an autonomous and controllable novel base editing platform is provided for crop functional gene research and molecular breeding.

The invention belongs to the field of gene engineering, and relates to a gramineous plant genome directional editing tool based on transposon-coded nuclease. The invention aims to solve the technical problem that the application range of a plant genome editing tool based on CRISPR-Cas9 and Cas12a is limited to a great extent due to the fact that CRISPR-Cas9 and Cas12a are large in protein size and difficult to deliver. The invention provides transposon nuclease suitable for genome editing of gramineous plants. The transposon nuclease is IsDge10, IsAam1 or enIscB subjected to codon preference optimization; and a gene editing system containing the three transposon nuclease is constructed. The gene editing system can be suitable for editing a coding region, a non-coding region and the like; the method can be widely applied to gramineous plants.

A method of decreasing conversion of nicotine to nornicotine is provided herein. The methods includes administering at least one basic region / leucine zipper (bZIP) type transcription factor inhibitor to an organism in need thereof. Also provided herein is a method of decreasing conversion of nicotine to nornicotine including mutating a bZIP type transcription factor binding site on a promoter of a nicotine N-demethylase (NND). Further provided herein is a method of decreasing conversion of nicotine to nornicotine including mutating a plant genome to knockout at least one bZIP type transcription factor.

The application discloses an anti-disease related protein OsMED16, a biomaterial thereof and application of the protein. The application solves the technical problem of improving plant disease resistance, reducing plant tillering number, delaying plant flowering time or preparing a plant with delayed flowering time. The application specifically provides a protein and a biomaterial thereof. The protein is any one of the following: A1) a protein with an amino acid sequence shown in sequence 2; A2) a protein with more than 80% identity with the protein shown in A1) and capable of improving plant disease resistance, which is obtained by substitution, deletion and / or addition of amino acid residues of the protein in A1); and A3) a fusion protein obtained by connecting a protein tag to the N terminal or / and C terminal of A1) or A2). Replacing a coding gene of the protein in a related gene in a plant genome can improve plant disease resistance, reduce plant tillering number and delay plant flowering time, and is used for rice variety improvement or breeding.

A non-naturally occurring heterologous CRISPR-Cas12a genomic editing system is described, including or encoding at least one Cas12a ortholog endonuclease selected from among Lb5Cas12a, CMaCas12a, BsCas12a, BoCas12a, MlCas12a, Mb2Cas12a, MbCas12a TsCas12a, and MAD7® ErCas12a endonucleases. The ortholog endonucleases enable genomic editing of plants at low temperatures, e.g., below 25° C. The CRISPR-Cas12a system can be employed for targeting PAM sites such as TTN, TTV, TTTV, NTTV, TATV, TATG, TATA, YTTN, GTTA, and GTTC, utilizing corresponding gRNAs, and is readily adapted to multiplexed editing operations of plants such as rice and other food crops.

The present application belongs to the field of genetic engineering. Specifically, the present application relates to a method for site-specific insertion of exogenous sequences in a genome. Specifically, the present application is based on a prime editing system (PE) using two adjacent pegRNAs with partially overlapping sequences on a reverse transcription template to achieve efficient and precise site-specific insertion of exogenous sequences in a genome, particularly a plant genome. Further coupling the system with a recombinase system such as Cre / Lox or FLP / FRT, etc. achieves site-specific insertion of large fragments of exogenous sequences in a genome, particularly a plant genome.

Provided is a simple and efficient method of plant genome editing that does not involve the incorporation of a gene cassette for the expression system of a site-specific DNA modification protein. The method of the present invention comprises: (i) a step for mixing plant cells or a tissue containing plant cells together with a site-specific DNA modification protein and a needle-shaped inorganic compound in a liquid medium and making a disturbance to perforate the cells with the needle-shaped inorganic compound, thereby introducing the site-specific DNA modification protein into the cells; and (ii) a step for culturing the plant cells or the tissue containing the plant cells, into which the site-specific DNA modification protein has been introduced in step (i), thereby causing a DNA mutation specific to a target site in the genome of the plant cells.

The present invention relates to a method for producing a MAGEA4-Fc transgenic plant, and can stably express the recombinant protein MAGEA4-Fc in which a human IgG Fc fragment is fused to a MAGEA4 protein through a platform using a tobacco plant. The method according to the present invention enables high-level expression efficiency and stable protein production by introducing a target gene into a plant genome by using an Agrobacterium binary vector. Mass production can be easily achieved by directly separating and purifying the recombinant protein from the transformed tobacco plant, and greater economic feasibility and expandability are provided compared to existing microbial or animal cell-based expression systems, thus providing greater utility as a fusion protein production platform.

The application belongs to the technical field of genetic engineering, and particularly relates to a negative-sense RNA virus plant genome editing system and application thereof. The genome editing system uses a CRISPR-Cas9 gene editing technology, takes a plant cytoplasmic rhabdovirus as a delivery carrier, is transmitted to a plant by means of a vector, utilizes an RNA movement motif, delivers gene editing components to an axillary meristem, and obtains mutant lateral buds along with plant tillering. The application can complete plant gene editing without tissue culture, overcomes plant tissue culture dependence, expands the range of gene editing lines, and only needs one generation to obtain a pure mutant without going through hybridization and selfing processes.

The invention discloses an editor fusing 5 'exonuclease and CRISPR-Cas12i2 as well as construction and application of the editor, and relates to the technical field of plant biology. The editor for fusing the 5'exonuclease and the CRISPR-Cas12i2 comprises a 5 'exonuclease T5E and Cas12i2 variant fusion protein T5E-Cas12i2 v1 coded by a nucleotide sequence, and a nuclear localization signal bpNLS, the 5' exonuclease T5E, the Cas12i2 variant Cas12i2 v1 and a nuclear localization signal bpNLS are sequentially arranged from the N end to the C end. Compared with an existing plant genome editing tool, the constructed T5E-Cas12i2v1 editor has the advantages that the efficiency is high, TTN-PAM sites are targeted, and site-specific large fragment deletion can be generated. The method is beneficial to accurate manipulation research on large cis-genetic elements, and has a wide application prospect in plant functional genome research and genetic improvement of crops.

The present invention relates to a method for producing a MAGEA3-Fc transgenic plant, and enables, through a platform using a tobacco plant, stable expression of a recombinant protein MAGEA3-Fc in which a human IgG Fc fragment is fused to a MAGEA3 protein. The method of the present invention enables high-level expression efficiency and stable protein production by using an Agrobacterium binary vector to introduce a target gene into a plant genome. The present invention enables recombinant proteins to be readily produced on a large scale through direct isolation and purification from transgenic tobacco plants, and is more economical and scalable than conventional microbial or animal cell-based expression systems, and thus can be effectively used as a fusion protein production platform.

Addressing the challenges of low genome editing efficiency and low efficiency in producing plants with mutations introduced into target genes, this research aims to provide plant genome editing technology with higher genome editing efficiency. [Solution] A method for producing genome-edited plants, comprising introducing a histone acetyltransferase expression cassette and a site-specific nuclease expression cassette into a plant.

The application belongs to the technical field of plant gene editing, and particularly relates to a plant genome editing system based on glycosylase MPG_V6.3 and application thereof. The application aims to develop a genome editing tool with high-efficiency base conversion and polynucleotide deletion. The technical scheme of the application is a coding gene of glycosylase MPG_V6.3, which has a nucleotide sequence as shown in Seq ID No. 1, 2065-2955. The application further provides a plant genome editing system based on the glycosylase MPG_V6.3, which comprises an MPG_V6.3-nCas9 fusion expression unit with a structure of promoter-MPG_V6.3-nCas9-terminator. The editing system realizes a leap from "single base editing" to "integrated editing of base editing and multi-base deletion", and can simultaneously realize base type conversion and multi-base deletion at a target site.

The invention provides a method for improving plant genome editing efficiency based on a transcription factor AtSOG1 and application of the method, and belongs to the technical field of gene editing. The invention finds that the AtSOG1 transcription factor has multiple influences on gene editing, a target plant is co-transformed by using an expression vector constructed by the AtSOG1 transcription factor and a gene editing vector, or an expression element of the AtSOG1 transcription factor is constructed into the gene editing vector, and then the target plant is transformed, so that the plant genome editing efficiency can be improved. The method provided by the invention not only can promote targeted gene knock-in based on a PE editor and HDR mediation, but also can improve the knockout efficiency of long fragments. Meanwhile, in the embodiment of the invention, an experiment is further designed, it is found that the AtSOG1 can promote a T-DNA unconformity knock-in event, and a T-DNA free knock-in plant is directly obtained in a T0 generation.

The invention discloses a plant genome prediction method, product and equipment based on a multi-dimensional data selection index, and belongs to the technical field of plant breeding. The method comprises the following steps: S1, data preparation: acquiring species data of a target species, and ensuring that multiple omics data of each individual are in one-to-one correspondence with phenotypes; s2, model construction: carrying out model training by adopting five-fold cross validation comprising two cycles of an inner layer and an outer layer so as to solve the problems of overfitting and generalization ability evaluation; s3, weight estimation: obtaining weight vectors of different omics based on a selection index strategy; s4, prediction calculation: using training set data to obtain phenotype prediction values of the prediction set by each group, assigning corresponding weights to obtain selection indexes combined with all group data, and repeating for a plurality of times to cover the whole data set; and S5, accuracy evaluation: calculating a decision coefficient of a selection index and a true value, and verifying a model effect by adopting a plurality of methods. According to the invention, the prediction precision of multiple omics can be obviously improved.

The invention discloses a plant genome nucleic acid fragment accurate insertion and plant in-vivo excellent nucleic acid screening method, and belongs to the technical field of biological gene engineering. The method comprises the following steps: (1) inserting an attb site into a plant genome, and carrying out genetic transformation on a plant to obtain a stably transformed plant; (2) constructing another transgenic vector containing a serine recombinase expression cassette, attp and donor donor fragments; and (3) delivering the transgenic vector into the plant cells in the step (1) through an agrobacterium tumefaciens-mediated plant genetic method, so that a fragment between the two attp sites on the T-DNA is accurately inserted into the plant genome. And carrying out character analysis and sequencing on the recombinant strain so as to obtain nucleic acid information associated with excellent characters of the plant. According to the method, the nucleic acid can be directionally inserted into a specific and consistent position, so that the result interference caused by the difference between the insertion position and the copy number is avoided, a researcher can accurately judge whether the plant character change is caused by the inserted nucleic acid or not, and the accuracy of a screening result is greatly improved.

The present disclosure provides a series of mutant Agrobacterium strains generated by random mutagenesis of a wide-type or ω mutant VirD2 gene or VirD2 protein. The mutant Agrobacterium strains of the present disclosure transiently express T-DNA-encoded transgenes in a target plant but do not stably integrate these genes into the plant genome.

The application belongs to the technical field of gene editing, and particularly relates to a Cas9 protein mutant and application thereof in plant gene editing. The application discloses a Cas9 protein mutant, an amino acid sequence of which is shown as SEQ ID NO. 1. The Cas9 protein mutant can efficiently screen cutting of various target sites, and is a Cas9 protein not dependent on NGG target site cutting. The application not only widens the targeting range of the Cas9 protein, but also provides the Cas9 protein mutant with higher gene editing efficiency, and has important application value in plant genome editing.

The application discloses a SYBR Green fluorescent quantitative PCR method for identifying Cyperus esculentus components based on a Ce44 gene and application thereof, and establishes a specific and high-sensitivity Cyperus esculentus component detection method by using a single copy gene of a Cyperus esculentus plant genome as a marker gene based on a SYBR Green real-time fluorescent quantitative technology.The method can accurately distinguish the Cyperus esculentus from four Cyperaceae plants and other 12 representative plants, and can accurately determine whether the six food samples on the market contain the Cyperus esculentus component.The specific real-time fluorescent quantitative PCR detection method for the Cyperus esculentus established by the application can provide a technical reference for qualitative and quantitative detection of the Cyperus esculentus component in Cyperus esculentus products.

The invention discloses a wheat TaWRKY65-3A gene high-yield excellent allelic variation detection method and application, and relates to the technical field of wheat genetic engineering.The method comprises the following specific steps that S1, genome DNA of a to-be-detected wheat material is extracted through a CTAB method or a plant genome DNA extraction kit, and the extracted genome DNA meets the PCR amplification requirement; s2, aiming at the 1265th SNP site of the TaWRKY65-3A gene, performing two rounds of PCR (Polymerase Chain Reaction) amplification by taking genome DNA as a template, and performing enzyme digestion treatment on a product of the two rounds of PCR amplification by using restriction enzyme Sma I; six key SNP sites are accurately determined by deeply analyzing the polymorphic sites of the TaWRKY65-3A gene, it is found that the sites have a linkage phenomenon in a wheat natural population, three main haplotypes are formed, allelic variation corresponding to the Hap-3A-2 haplotype is high-yield and excellent allelic variation, and on the basis of the key SNP sites, especially the 1265th SNP site and the 1658th SNP site, the high-yield and excellent allelic variation is obtained. A specific primer and a dCAPS marker are designed, and a complete detection technical scheme is established.

Transgenic INIR12 maize plants comprising a vip3Aa19 or vip3Aa20 expression cassette linked to a secondary nopaline synthase terminator element which lack a selectable marker gene and / or which comprise modifications that provide for facile excision of the INIR12 transgenic locus from the maize plant genome are provided. Genomic DNA of INIR12 transgenic plants, detection of INIR12 plants and products thereof, methods of making INIR12 plants, and use of INIR12 plants to facilitate breeding are disclosed.

The invention discloses an improved guide editor and application thereof, and relates to the technical field of gene editing. Comprising Ec48 reverse transcriptase or a variant thereof, Tf1 reverse transcriptase or a variant thereof or M-MLV reverse transcriptase or a variant thereof; or, a first fusion protein, the first fusion protein comprising: a first reverse transcriptase and a second transcriptase; wherein the first reverse transcriptase or the second reverse transcriptase is selected from Ec48 reverse transcriptase or a variant thereof, Tf1 reverse transcriptase or a variant thereof, or M-MLV reverse transcriptase or a variant thereof. The improved guide editor can realize accurate editing in the plant genome, and the editing efficiency is high.