129 results about "Start codon" patented technology

To provide a medicine for treating a disease in a mammal.SOLUTION: Provided is a medicine for treating a disease in a mammal, comprising an expression cassette that is a first expression cassette comprising a chimeric gene operably linked to a first expression control element, the chimeric gene comprising a first portion comprising a minigene having an alternatively spliced exon and a second portion encoding a first RNA encoding a protein, the first portion being 5' to the second portion, the second portion lacking an initiation codon or start codon, the alternatively spliced exon comprising a translation initiation regulatory sequence, expression of the protein being controlled by inclusion of the alternatively spliced exon, and the medicine being used in combination with a small molecule splicing modifier that modulates splicing of the alternatively spliced exon.SELECTED DRAWING: Figure 1

The present application relates to a kind of deletion mutant nucleic acid and its application in anti-herbicide.The deletion mutation occurs in the promoter of rice OsHPPD Gene, the length of deletion sequence is at least 10 bp, and the A in the start codon ATG of gene is 0, at least the bases located in the upstream of start codon ATG from 2041 to 2032 are deleted. OsHPPD The present application finds that by deleting part of the sequence in the promoter of rice OsHPPD Gene, rice can obtain resistance to HPPD inhibitor herbicides, which is of great application value for ensuring agricultural production safety and improving the efficiency of herbicide use.

The invention provides an SNP (Single Nucleotide Polymorphism) site of an ABO blood group variant, and a B subtype variant blood group identification kit and method, and relates to the technical field of biology. The SNP (Single Nucleotide Polymorphism) sites of the ABO blood group variant comprise the 842th c.842Cgt from an initiation codon of an ABO * B.01 allele coding region of an ABO gene; mutation of T. A substance for detecting the SNP site of the ABO blood group variant can be applied to preparation of a B subtype variant blood group identification method and product, so that the substance can be effectively applied to rapid detection of patients and blood donors, the B subtype variant blood group of an individual can be defined, and potential possible blood transfusion reaction can be prevented in time.

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 present invention provides a method for computer analysis of a genome of a plant subject, comprising identifying a plurality of sequences that collectively define a promoter set of the plant subject, where each sequence within the promoter set consists of an expression control region, the expression control region extends from an initiation codon of an open reading frame to about 100 kilobases away from the 5'and / or 3 'end of the initiation codon. A transcriptome in the form of mRNA expression data of a plant subject is obtained and analyzed using a sequence-based modeling algorithm to provide a predicted value of the expression level of each protein coding gene within the transcriptome and associate this value with the sequence of the corresponding expression control region within the promoter group. The method can be used to generate new designs for expression control sequences, which are most likely to provide the desired expression profile for operably linked coding sequences. Nucleic acid libraries, plant cells, plant tissues and plants comprising the novel expression control sequences are also provided.

The invention discloses a 5 'UTR library, a recombinant expression vector and a genetically engineered bacterium for improving the yield of folamin in pathogenic bacterium Badei and application of the 5' UTR library and the genetically engineered bacterium, and belongs to the technical field of microbial genetic engineering. The method comprises the following steps: designing an SD region based on a 16S rRNA (ribosomal Ribonucleic Acid) 3'end complementary sequence of a Buddei pathogenic bacterium XBD101 serving as an original strain, regulating and controlling the number of bases between the SD sequence and an initiation codon AUG and optimizing an UTR secondary structure to construct a library containing 14 UTR sequences, and replacing three sequences N4A, N7A and N18A with the strongest translation ability in the library to the UTR region of a folamin synthesis key gene fclC, so as to obtain the folamin peptide. And finally, the capacity upgrading of the folamin chassis strain is realized, and an important support is provided for the industrial production of agricultural antibiotics.

The present invention is a capped RNA which has a translated region beginning with a start codon and encoding a protein and an upstream region upstream of the start codon, and a branch part having a cap structure. This branch part preferably has at least one structure of the following (a) to (c): (a) a capped probe having a sequence complementary to a sequence of a part or all of the untranslated region and having the cap structure is hybridized to the untranslated region; (b) a branched chain branching from the middle of a main chain constituting the RNA is provided in a part of the untranslated region and the cap structure is provided in the branched chain; and (c) the RNA has a branch structure in which the 51 end of a main chain of the untranslated region is branched into two or more branches, the cap structure being provided in each branch.

The invention relates to a KASP molecular marker of a wheat quality related gene TaSF3B4 and application of the KASP molecular marker. The KASP molecular marker is KASP-TaSF3B4, and an SNP (Single Nucleotide Polymorphism) site (A / G) is detected at 248421161bp of a wheat 3B chromosome, namely, the distance between a TaSF3B4 promoter region and an initiation codon is 1127bp. According to the marker, a TaSF3B4 gene influencing the wheat quality character is obtained according to a way that differentially expressed genes in 15d transcriptome data of a grain extreme material are extremely remarkably enriched to a spliceosome, and regular SNP loci exist through sequence comparison and analysis of the TaSF3B4 genes in different varieties and upstream and downstream sequences of the TaSF3B4 genes. The functional KASP molecular marker of the wheat poly (adenylic acid binding protein) gene TaSF3B4 provided by the invention can be used for identifying whether TaSF3B4 excellent alleles exist in wheat varieties / strains or not, and can be applied to assisted selective breeding and pyramiding breeding of other genes related to known quality.

A method for predicting translation efficiency of an mRNA vaccine comprises the steps in which: an analysis device receives sequence information of a target mRNA vaccine; the analysis device extracts, from the sequence information, an input sequence including a 25 nt sequence to which a primer binds in a 5' untranslated region (UTR), a 50 nt sequence of the 5' UTR immediately before a coding sequence (CDS) site, and a 30 nt sequence after a start codon of a coding region; the analysis device calculates secondary structure information for the input sequence; and the analysis device inputs the input sequence and the secondary structure information into a pre-trained deep learning model so as to predict the translation efficiency of the corresponding mRNA vaccine.

The invention discloses an mRNA sequence scoring method and related equipment. The method comprises the following steps: acquiring an mRNA sequence to be scored; extracting a key base sequence from the mRNA sequence, wherein the key base sequence comprises an initiation codon context and a termination codon context; inputting the key base sequence into a target scoring model, and processing to obtain a score of the mRNA sequence in the target index; wherein the target scoring model comprises a multi-modal RNA sequence coding module and a large language model, and the target scoring model is obtained based on unsupervised pre-training of transcription sample data and supervised fine tuning of a score label data set corresponding to the target index. According to the method, a non-coding region and a coding region key region (start / stop codon context) are integrated as input, and a large language model trained based on mass data is used for processing, so that various complex factors such as sequence context information and secondary structures can be comprehensively considered, and the method can be widely applied to the technical field of gene sequence analysis.

The present invention discloses a method for constructing acid-resistant engineered algae, belonging to the fields of biology and molecular biology. The specific method is as follows: Using Phatr3_J33543 with the sequence structure shown in SEQ ID No.1, Phatr3_J50516 with the sequence structure shown in SEQ ID No.2, or Phatr3_Jdraft1806 with the sequence structure shown in SEQ ID No.3 as the target gene, removing the stop codon of the target gene, and connecting the fluorescent protein gene without the start codon through the DNA sequence encoding glycine to obtain a recombinant gene; inserting the recombinant gene into the multiple cloning site of the pPhaNR plasmid to obtain a vector plasmid; introducing the vector plasmid into diatom cells by electroporation and then inoculating them onto the ESAW solid selection medium containing bleomycin, and growing brown colonies; selecting the colonies with fluorescent protein signals in the colonies, which are the acid-resistant engineered algae. Compared with the wild algal species, the acid-resistant ability of the engineered algae obtained by the present invention is significantly improved. The present invention has important reference significance for the cultivation of acid-resistant diatoms.

The application discloses a method for quantitatively regulating gene translation and application thereof. The method comprises the following steps: sequencing translation efficiency of Kozak sequences and variants thereof before a start codon of a target gene or before an upstream open reading frame of a non-coding region of the target gene, and performing gene editing on the Kozak sequences according to a sequencing result, so as to realize in-situ manipulation of the Kozak sequences and quantitatively regulate gene translation, and the Kozak sequences and the variants thereof are 3-6 bp in length respectively. The application establishes a highly efficient, flexible and widely applicable gene expression regulation method. By using a precise gene editing tool to customize the Kozak sequences of the target gene in-situ, the expression level of the target gene can be quantitatively controlled at the gene translation stage.

The present invention is a capped RNA which has a translation region starting from an initiation codon and encoding a protein, and an upstream region on the upstream side of the initiation codon, and which comprises a branch having a cap structure. It is preferable that the branch section has at least one of the following structures (a) to (c). (a) a capped probe having a sequence complementary to some or all of the sequences of the untranslated region and having a cap structure hybridizes to the untranslated region; (b) a branch that branches from the midway of the main chain constituting the RNA is provided in a part of the untranslated region, and a cap structure is provided in the branch; (c) The 5'end of the main chain of the untranslated region has a branch structure divided into two or more branches, and each branch is provided with a cap structure.

A bioengineering method which comprises introducing an inert nucleic acid cassette into a biological entity without introducing or modifying characteristics or traits in the biological entity. The method comprises receiving or providing a sample comprising the biological entity having a nucleic acid sequence; selecting an integration site in the nucleic acid sequence for inserting the inert nucleic acid cassette; designing the inert cassette with optimized primer sequences, optimized probe sequences, optimized stop codons and disrupted start codons, and inserting the inert nucleic acid cassette into the biological entity at the integration site; and validating that no characteristics have been added or modified in the biological entity.

The invention provides a 5 'UTR (Untranslated Region) element for improving protein expression quantity and application of the 5' UTR element, and belongs to the technical field of biology. The sequence of the 5 'UTR element is shown as SEQ ID NO.1. Modified mRNA is obtained by adding the 5' UTR element between an initiation codon of unmodified mRNA and 5 'UTR. Compared with the unmodified mRNA, the target protein expression quantity of the modified mRNA is remarkably improved, and meanwhile, the 5' UTR element is good in universality and suitable for improving the expression quantity of most mRNA, so that the 5 'UTR element has a good application prospect.

A microbial gene editing scheme automatic design method, system and storage medium; the method comprises: selecting a design object for different types of strains, the design object is bacteria or fungi; after determining the target gene, obtaining the target gene information from the NCBI database; the design scheme of bacteria comprises the following steps: determining the gRNA design region; selecting the optimal gRNA sequence; doing sequence complexity and GC content analysis; the design scheme of fungi uses a frameshift scheme: the gRNA of the frameshift scheme is set at any position not overlapping with other genes after the start codon in the gene; select gRNA sequence in gRNA design region, select the optimal gRNA sequence; design two pairs of four primer sequences for homologous arm identification primer; this scheme can replace the complicated and time-consuming manual scheme, save a lot of time, solve the problem of high cost in the design of the best site and the design of sgRNA sequence scheme of artificial gene knockout.

The invention relates to the technical field of plant breeding, in particular to a mutant of an FZP gene promoter and application of the mutant. Compared with a wild type rice FZP gene promoter, the mutant has any one of the following mutations: (1) 5bp is deleted from 3995bp to 3989bp at the upstream of an initiation codon; and (2) inserting 1bp between 3991bp and 3990bp at the upstream of the initiation codon. The application comprises the following steps: i) increasing the grain density, the spikelet number per ear, the secondary branch number or the yield of the oryza plants; ii) cultivating a transgenic Oryza plant; the FZP gene promoter mutant has the advantages that the FZP gene promoter mutant can be used for regulating and controlling the stem differentiation process of rice, promoting the increase of the grain density of the rice and increasing the yield of the rice, and the FZP gene promoter mutant has an important application value.

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.

The application relates to the technical field of wheat kernel type genes, and provides a wheat kernel type gene TaCYT-5A TaCYT-5A The application of the molecular marker is KASP-TaCYT-5A, the marker is a SNP site (C / T) detected at 495818646 bp in a QTL cluster for controlling the wheat kernel type and kernel length traits of a 5A chromosome 484432128-495883712 bp, that is, a 4-hydroxyphenylacetaldehyde oxime monooxygenase gene TaCYT-5A The coding region is 124 bp away from the start codon. The functional KASP molecular marker of the wheat 4-hydroxyphenylacetaldehyde oxime monooxygenase gene TaCYT-5A provided by the application can be used for identifying whether an TaCYT-5A excellent allele exists in a wheat variety / strain, and applied in assisted selection breeding and breeding in combination with other known kernel type related genes.

The invention relates to the technical field of gene editing, and particularly provides a method for creating fluazifop-p-butyl herbicide resistant rice through gene editing of an OsACCase gene promoter region. The method comprises the following steps: constructing a crRNA plasmid library covering an OsACCase gene promoter region on the basis of a CRISPR / LbCas12a system, performing agrobacterium-mediated transformation on rice callus, and performing resistance screening, fluazifop-p-butyl herbicide screening and molecular identification to obtain a fluazifop-p-butyl resistant rice strain. The strain is subjected to-4bp / -5bp double allelic deletion mutation in an upstream 1865-site region of an initiation codon ATG of the OsACCase gene, and a homozygous mutant strain and a double allelic deletion mutant strain can normally grow after being sprayed with 6 times of field recommended dose of fluazifop-p-butyl. The herbicide resistance is realized by editing the gene promoter region, the growth defect of coding region mutation is avoided, the breeding technology is enriched, and weed prevention and control are assisted.

A toehold exchange riboregulator gate includes: a gate input toehold domain; a ribosome binding sequence; a start codon; a c spacer domain positioned between the ribosome binding sequence and the start codon; a protein coding sequence; and a self-cleaving ribozyme sequence that produces a double-stranded RNA gate suitable for strand exchange and produces the gate input toehold domain.

The invention relates to the technical field of gene engineering, in particular to a mutant gene and protein for causing rice flag leaves to become smaller and application of the mutant gene and protein. According to the mutant gene capable of causing the rice flag leaf to become smaller, the nucleotide sequence of the mutant gene is a nucleotide sequence shown as SEQ ID NO.1 inserted into the 153rd site of the nucleotide sequence of a wild rice OsGATA15 gene from an initiation codon ATG. According to the invention, an indica type rice line 93-11 is subjected to < 60 > Co-gamma radiation treatment to obtain a new flag-leaf-reduced mutant ym66, and the ym66 mutant is hybridized with the indica type rice line YHSM to obtain an excellent flag-leaf-reduced restorer line NP27. In the seed production process, the seed production yield is not reduced under the condition that flag leaves of NP27 are not cut, and in addition, the F1 generation shows excellent characters in the aspects of plant type and thousand seed weight.

The application discloses an mRNA sequence scoring method and related equipment, and the method comprises the following steps: acquiring an mRNA sequence to be scored; extracting a key base sequence from the mRNA sequence, wherein the key base sequence comprises a start codon context and a stop codon context; inputting the key base sequence into a target scoring model to obtain a score of the mRNA sequence on a target index; wherein the target scoring model comprises a multi-modal RNA sequence encoding module and a large language model, and the target scoring model is obtained based on unsupervised pre-training of transcription sample data and supervised fine-tuning of a score label data set corresponding to the target index. The application integrates non-coding regions and key regions of coding regions (start / stop codon contexts) as inputs, and processes them by using a large language model trained based on massive data, so that the application can comprehensively consider various complex factors such as sequence context information and secondary structure, and can be widely applied to the technical field of gene sequence analysis.

Provided herein are RNA editing oligonucleotides capable of effecting ADAR-mediated deamination of a target adenosine comprised in a target RNA, wherein the oligonucleotide comprises a sequence that is capable of hybridizing with a region in the target RNA comprising said target adenosine, and wherein the oligonucleotide comprises at least one conformationally restricted nucleotide (CRN) positioned at the 5' and / or at the 3' terminus of the oligonucleotide and / or internally. In particular, the target adenosine may be part of a therapeutic target codon in the MECP2 protein-coding RNA. Provided herein are RNA editing oligonucleotides capable of effecting ADAR-mediated deamination of a target adenosine in a target RNA molecule encoding a target protein, wherein the oligonucleotide comprises a sequence that is capable of hybridizing with a region in the target RNA molecule comprising said target adenosine, and wherein the target adenosine is part of a codon encoding a posttranslational modification site in the target protein. In particular, the target adenosine may be part of a ubiquitination site in the frataxin (FXN) protein. Provided herein are RNA editing oligonucleotides capable of effecting ADAR-mediated deamination of a target adenosine in a target RNA molecule encoding a target protein, wherein the oligonucleotide comprises a sequence that is capable of hybridizing with a region in the target RNA molecule comprising said target adenosine, and wherein the target adenosine is part of a start codon in the 5' untranslated region of the target RNA molecule. In particular, the target adenosine may be part of the start codon of an untranslated ORF (uORF) in the human Sodium-dependent phosphate transporter 2 protein (SLC20A2).

The invention relates to the technical field of gene editing, and particularly provides rice OsEPSPS gene promoter region deletion mutation and application thereof in creation of herbicide-resistant rice, the deletion mutation is that a sequence with the length of 22bp or 28bp is deleted in the OsEPSPS gene promoter region, namely, 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; and deleting the basic groups at sites 2534 to 2555 upstream of the initiation codon ATG or the basic groups at sites 2527 to 2554 upstream of the initiation codon ATG. According to the application disclosed by the invention, the rice can obtain resistance to EPSPS inhibitor herbicides by deleting part of sequences in the promoter of the rice OsEPSPS gene, so that the application has great application values in guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

The invention relates to a rice OsACC gene promoter mutant for resisting ACCase inhibitor herbicides and application of the rice OsACC gene promoter mutant. The rice OsACC gene mutant is a deletion mutant or a deletion and combination insertion mutant; the deletion mutation occurs in the promoter of the rice OsACC gene, and the sequence of the nucleic acid before deletion mutation is as shown in SEQ ID No. 1; the length of the sequence of the deleted nucleic acid is 29 bp to 97 bp, A in the initiation codon ATG of the OsACC gene is 0 site, the deleted nucleic acid occurs in the 305-209 sites of the upstream of the initiation codon ATG, and at least the 271-243 sites of the upstream of the initiation codon ATG are deleted; the inserted nucleic acid occurs within sites 271 to 243 upstream of the initiation codon ATG. The mutant can enable rice to obtain resistance to at least one of herbicides such as haloxyfop-R-methyl, fluazifop-p-butyl and clethodim.

The present invention discloses a molecular marker, primer and application thereof for identifying the color of Actinidia arguta fruit. The molecular marker of the present invention is located at 495bp upstream of the start codon of the AaLDOX gene promoter region of Actinidia arguta, which is a 29bp Indel variation. Utilizing the molecular marker and method of the present invention, the fruit color can be quickly and accurately identified in the childhood stage of Actinidia arguta by conventional molecular test detection, and the red and green Actinidia arguta germplasm resources can be efficiently distinguished. The method can be used for parent selection and seedling identification of offspring, greatly improving the efficiency and accuracy of selective breeding. The method of the present invention can be detected in each tissue, organ and each developmental stage of the plant, and has the advantages of being convenient and fast, having good specificity, high accuracy, and being unaffected by climate and environment. It can be applied to the precise, directional and efficient breeding of red Actinidia arguta, and has a large application value.

The invention features lentiviral transfer vectors that include heterologous nucleic acids to be introduced into a cell. The lentiviral transfer vector may be characterized by the following features: (a) including a cytomegalovirus (CMV) promoter; (b) including a polynucleotide encoding a partial gag protein that includes a mutated INS1 inhibitory sequence that reduces restriction of nuclear export of RNA; (c) not including a polynucleotide encoding the INS2, INS3, and INS4 inhibitory sequences of gag; (d) not including an SV40 origin of replication and / or an f1 origin of replication; (e) including a cPPT sequence that contains splice site; (f) including an EF1 alpha promoter with intact splice donor and acceptor sites; and (g) including hepatitis B PRE with mutation in start codon of X protein ORF.

The invention discloses application of a rice OsME5 gene and a molecular marker thereof in regulation and control of rice dry direct seeding seedling emergence, the nucleotide sequence of the rice OsME5 gene is as shown in SEQ ID No.1, and the coding sequence of OsME5 protein coded by the rice gene OsME5 is as shown in SEQ ID No.2; the method comprises the following steps: knocking out an OsME5 gene by adopting a CRISPR / Cas9 method, and designing a molecular marker by utilizing natural nucleotide polymorphism (SNP) of an OsME5 promoter region SEQ ID No: 3; the OsME5 has a key SNP OsME5-578G / A at 578 base pairs on the upstream of an initiation codon, the seedling emergence ability of the germplasm carrying the OsME5-578G is relatively strong when the germplasm carrying the OsME5-578G is covered with soil and the seedling emergence ability of the germplasm carrying the OsME5-578A is relatively weak when the germplasm carrying the OsME5-578G is covered with The OsME5 gene disclosed by the invention can be applied to the field of agriculture, and a candidate gene is provided for improving the seedling emergence capability of rice soil-covering dry direct seeding.

The invention provides a preparation method and application of a non-human mammal model of hyperuricemia. Specifically, the invention provides application of the gene editing reagent and preparation of the kit for constructing the non-human mammal model of hyperuricemia, the gene editing reagent comprises a reagent for introducing an additional second initiation codon ATG at the upstream of an initiation codon ATG of a UOX gene, and when the second initiation codon ATG is introduced, the wild type mRNA transcript level and / or the wild type UOX protein expression level of the UOX gene are / is reduced. The animal model provided by the invention is an effective animal model for simulating human hyperuricemia, and has the advantages of no renal toxicity and longer survival time. The method can be used for long-term stable research of related diseases such as hyperuricemia and can be used for screening and testing of specific drugs.