97 results about "Translational efficiency" patented technology

The invention discloses an mRNA non-coding region sequence optimization method and system and electronic equipment, and the mRNA non-coding region sequence optimization method comprises the steps: constructing an initial candidate library according to a target protein; inputting the initial candidate library into a pre-trained mRNA sequence optimization model to obtain a prediction data set; performing multi-dimensional scoring and sequence optimization on the prediction data set to obtain a sequence recommendation group; performing biological verification on the sequence recommendation group to obtain an optimized mRNA sequence; wherein the prediction data set comprises a sequence ID, a sequence content, a prediction TE score and a confidence interval. According to the method, the translation efficiency of the mRNA sequence can be efficiently and accurately predicted, the candidate sequence with high expression potential is screened out, meanwhile, the consumption of computing resources is reduced, and the overall design cost is reduced.

The invention provides a method for predicting mRNA translation efficiency and a prediction system. The method comprises the following steps: acquiring mRNA sequence data, adding a classification mark at the starting end of the mRNA sequence data, and processing the mRNA sequence data with the classification mark to obtain an embedded sequence; extracting local features by using a first feature extraction module, extracting global dependency features by using a second feature extraction module, extracting time sequence features by using a third feature extraction module, and extracting external features by using a fourth feature extraction module; modulating the local features by using the global dependency features to obtain modulated local features; and obtaining fusion features based on the modulated local features, the time sequence features and the external features, and performing prediction based on the fusion features to obtain a prediction result of the mRNA translation efficiency. Therefore, the accuracy of predicting the mRNA translation efficiency can be improved.

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 invention discloses a 5 'UTR sequence and application thereof in improving mRNA translation efficiency, and belongs to the technical field of biology, the designed 5' UTR sequence has an excellent mRNA expression promoting function, and compared with a common HBA1 gene 5 'UTR, the FLuc mRNA expression level fused with the 5' UTR can be improved to 2-3.5 times.

The present invention relates to: a recombinant RNA molecule having improved expression efficiency of a target protein; and a DNA molecule encoding same. The present invention is based on the discovery of five 5' UTR sequences derived from F2, F9, CFHR3, MBL2, and C3 genes as optimal heterologous 5' UTR sequences that commonly increase protein expression with the greatest efficiency in various tissues in vivo, and these UTRs showed a protein expression increase equivalent to or up to 2 times greater than 5' UTR applied to mRNA vaccines that are currently available commercially. Accordingly, the present invention can be effectively used as an excellent nucleic acid therapeutic composition capable of stably and systemically producing a therapeutically effective amount of a pharmacological protein in the body of a patient.

The invention discloses base-modified mRNA (messenger Ribonucleic Acid) and application thereof in a cell-free protein synthesis system, and belongs to the technical field of nucleic acid modification. Natural nucleoside triphosphate is completely replaced by different types of basic group modified nucleoside triphosphate, mRNA with different chemical modifications is obtained through an in-vitro transcription method and purification, and the mRNA is put into an in-vitro cell-free protein synthesis system for reaction. A reaction element recombination system-PURE system based on prokaryote Escherichia coli purified protein and an eukaryote CFPS system taking rabbit reticulated red lysate as a material are mainly selected to detect the corresponding translation efficiency of mRNA modified by different bases in two cell-free protein synthesis systems respectively. Different regulatory activities may be shown in different cell-free protein synthesis systems when different modified nucleotides are doped into mRNA, base modified triphosphoric acid capable of improving the translation efficiency of an in-vitro protein synthesis system is screened out, and the stability of mRNA in the cell-free protein synthesis system and the corresponding protein expression level are improved.

The invention discloses a recombinant full-length human-derived III-type collagen as well as a preparation method and application of the recombinant full-length human-derived III-type collagen. The method comprises the following steps: replacing at least one corresponding rare codon of glycine, proline, arginine, leucine, isoleucine, valine and / or glutamic acid host bacteria in a Gly-Pro-X or Gly-X-Pro tripeptide structure coded by a wild full-length human III-type collagen gene sequence with a synonymous codon used by the host bacteria at high frequency; the translation efficiency in prokaryotic systems such as escherichia coli is obviously improved; the method comprises the following steps: designing 6-12 gene segments with overlapping sequences, splicing by adopting an overlapping extension PCR (Polymerase Chain Reaction) technology to obtain a full-length optimized gene, constructing the full-length optimized gene to an expression vector, and transforming the full-length optimized gene into host bacteria for induced expression; the expression product exists in the form of an inclusion body, and the natural conformation of the expression product is recovered through in-vitro renaturation; according to the method, high-efficiency expression and active preparation of the full-length human-derived III-type collagen in a prokaryotic system are realized.

The invention discloses a polyA tail element, a construction body and application of the polyA tail element and the construction body. The polyA tail element is inserted into a sequence capable of mutually complementarily pairing basic groups behind A at any one of 27th to 33rd and 85th to 91st in the direction from 5'to 3 '; or the basic group at the multiple site of the 12th, 13th, 14th or 15th of the polyA tail element is C. The polyA tail element disclosed by the invention is high in correct cloning rate and good in passage stability, and can be used for fermentation production of mRNA template plasmids; a construct or a recombinant expression vector containing the gene has strong stability and high translation efficiency, and can efficiently express a target protein; and a new template plasmid can be quickly transformed and constructed for transcriptional production of new mRNA, so that the production time and cost are saved.

The application relates to an mRNA untranslated region translation efficiency prediction method and device, computer equipment and a storage medium. The method comprises the following steps: extracting sequence features and structure features of an mRNA sequence to be predicted, the structure features being generated by encoding based on at least one of free energy change and secondary structure of the mRNA sequence to be predicted; inputting the sequence features and the structure features into a trained translation efficiency prediction model for prediction to obtain a translation efficiency prediction result of an untranslated region in the mRNA sequence to be predicted. The method can improve the accuracy of the mRNA untranslated region translation efficiency prediction result.

The invention discloses a 5 'UTR sequence and application thereof in improving mRNA translation efficiency, and belongs to the technical field of biology, the designed 5' UTR sequence has an excellent mRNA expression promoting function, and compared with a common HBA1 gene 5 'UTR, the FLuc mRNA expression level fused with the 5' UTR can be improved to 2-3.5 times.

The invention discloses an mRNA (messenger Ribonucleic Acid) non-coding region sequence screening method and system and electronic equipment. The method comprises the following steps: constructing a candidate screening set; for each sequence in the candidate screening set, calculating related features of the RNA secondary structure to obtain corresponding feature vectors; based on a preset mRNA drug engineering constraint rule, filtering the candidate screening set to obtain a feasible candidate set; inputting the feasible candidate set and the feature vector into a pre-trained structure enhanced pre-training language model to obtain a performance prediction score of each candidate sequence; and performing descending sorting on the performance prediction score of each candidate sequence, and outputting the first k candidate sequences. According to the method, RNA secondary structure features can be explicitly fused into representation learning, compared with a pure sequence model, the root mean square error on a translation efficiency prediction task can be reduced by about 15%-25%, and the structure sensing ability is improved; through engineering constraint filtering, sequences which cannot be synthesized or have security risks can be systematically eliminated, and the constraint passing rate in Top-K output reaches 100%.

The invention discloses an application of an engineered mRNA (messenger Ribonucleic Acid) medicine for coding COL3A1 protein in resisting skin photoaging. According to the method, accurate rational design and optimization are carried out on the 5'untranslated region (UTR) of the hCOL3A1 mRNA, so that the translation efficiency and the protein yield of the hCOL3A1 mRNA are greatly improved, the expression level of the hCOL3A1 target protein far beyond the conventional sequence in vivo and in vitro is realized, and the type III collagen is efficiently and endogenously synthesized in skin cells. Experimental results show that the engineered mRNA can effectively reduce the skin oxidative stress level, inhibit cell aging and promote endogenous synthesis and deposition of corium layer collagen, so that the barrier function of photoaged skin is remarkably improved, the dermis thickness is increased, the tissue structure is repaired, and systemic toxicity or immune side effects are not observed. The invention provides a novel strategy with high efficiency and safety for resisting skin photoaging.

Provided are a translation regulatory element, an expression vector, a pharmaceutical composition, a vaccine composition, a construction method, a method for expressing a target protein, and the use thereof. The translation regulatory element is selected from recombinant IRES sequences having sequences as shown in SEQ ID NOs: 8-11, and / or one or more segments of recombinant IRES sequences complementary to the sequences as shown in SEQ ID NOs: 8-11; and can show high translation efficiency when being used for regulating gene expression, thereby realizing high-level expression of a target gene.

The present invention is related to method of preparing a gain-of-function mutation in poxvirus, like buffalopox virus (BPXV) by long-term propagating it in the presence of methyltransferase inhibitor like 3-Deazaneplanocin A (DZNep), which results in generation of viral 2'O-MTAse (VP39) mutant with a single point mutation (M236I). The resultant mutant has the following characteristics: (a) M236I mutation of 2'O-MTAse increases the cap-1 markings on viral mRNA; (b) The 2'O-MTAse mutant virus had hyperactive 2'O-MTAse and exhibited enhanced stability and translational efficiency of its mRNA; (c) The capping (cap1) performed by the mutant viral 2'O-MTAse (VP39) results in higher viral protein production, as compared to the wild type VP39 and (d) Mutant virus replicated at higher viral titer than the wild-type virus.

The application discloses a PACE structure modified cap analog, which is based on the existing cap analog structure, has ribose bases or modified ribose bases at both ends, three ribose structures in the middle and a triphosphonate segment or a phosphonate segment connecting the ribose structures, and has one or more PACE structures or PACE derivative structures on the triphosphonate segment or the phosphonate segment, namely, (alkoxy)alkyl-COOH or a salt thereof with or without substitution on the triphosphonate or the phosphonate, so that the IVT yield and capping efficiency are improved, and the translation efficiency of the target mRNA is improved.

The application provides a method and a prediction system for predicting mRNA translation efficiency. The method comprises obtaining mRNA sequence data and adding a classification mark at the start of the mRNA sequence data, processing the mRNA sequence data with the classification mark to obtain an embedded sequence; extracting local features using a first feature extraction module, extracting global dependency features using a second feature extraction module, extracting timing features using a third feature extraction module, and extracting external features using a fourth feature extraction module; modulating the local features using the global dependency features to obtain modulated local features; obtaining fusion features based on the modulated local features, the timing features, and the external features, and performing prediction based on the fusion features to obtain a prediction result of the mRNA translation efficiency. In this way, the accuracy of predicting the mRNA translation efficiency can be improved.

The invention provides a GLP-1-Fc fusion protein synthesis culture medium which is completely composed of chemical definition components, does not contain complex organic nitrogen sources such as yeast extract powder and peptone, and is mainly composed of a carbon source, an inorganic nitrogen source, vitamins, trace elements, inorganic salt and amino acid. On the basis of an inorganic nitrogen source, high-proportion essential amino acids in target proteins such as Val, Lys, Leu and Phe are directionally supplemented, and sodium citrate is added as a TCA cycle intermediate to strengthen energy metabolism. According to the strategy, the intracellular amino acid self-synthesis energy consumption can be remarkably reduced, and the translation efficiency and metabolic balance are improved; a 5L fermentation tank verification result shows that the final yield of the GLP-1-Fc fusion protein reaches 0.72 mg / mL and is more than two times that of a BSM culture medium, and impure proteins are remarkably reduced; the culture medium disclosed by the invention is controllable in formula, high in repeatability, capable of effectively reducing the metabolic burden and increasing the expression quantity of target protein, and suitable for industrial production of GLP-1 analogues and other high-amino-acid bias proteins.

The application belongs to the field of biotechnology and medicine, and more particularly relates to mRNA encoding red fluorescent protein and application thereof. The mRNA encoding red fluorescent protein comprises an ORF sequence of red fluorescent protein, and the nucleotide sequence is as shown in SEQ ID NO: 5 to 13. The mRNA encoding red fluorescent protein provided in the application has high translation efficiency by reasonably optimizing the nucleic acid sequence of the functional region, so that the output of red fluorescent protein is increased, and the mRNA encoding red fluorescent protein has wide application prospects in the field of gene drug development.

The present invention relates to artificial nucleic acid molecules comprising at least one open reading frame and at least one 3'-untranslated region element (3'-UTR element) and / or at least one 5'-untranslated region element (5'-UTR element), wherein said artificial nucleic acid molecule is characterized by a high translation efficiency. Said translation efficiency is at least partially due to said 5'-UTR element or said 3'-UTR element, or due to both said 5'-UTR element and said 3'-UTR element. The present invention further relates to the use of said artificial nucleic acid molecules in gene therapy and / or gene vaccination. Furthermore, novel 3'-UTR elements and 5'-UTR elements are provided.

This invention belongs to the fields of genetic engineering and biomedicine, specifically relating to a method for codon optimization of human coagulation factor IX and the recombinant human coagulation factor IX encoding gene and its applications. Based on RiboDecode, this invention optimizes relevant parameters to improve the codons of the target gene, thereby increasing the expression level and activity of the target gene in the host. The optimized FIX gene mRNA molecule exhibits good stability and high translation efficiency; further addition of GCCACC upstream of the start codon further enhances expression efficiency. This invention optimizes transfection-related operations, identifies optimal transfection conditions, and achieves maximum expression levels through transient transfection, effectively improving the yield of recombinant human coagulation factor IX. The results of the examples show that the yield of recombinant human coagulation factor IX can reach 1.58±0.10 mg / L, and the coagulation activity can reach 66.8±0.51%, with significant optimization of all key indicators.

A linearized mRNA preparation system, use thereof, and a preparation method for preparing mRNA by using same. A linearization buffer system comprises: 0-150 mM Tris-Ac or phosphate, 0.5-150 mM divalent metal ions or trivalent metal ions, 0.1-0.75 U / μL restriction endonuclease, and 0-7.5 mM spermidine. The present invention provides a transcription system, which has higher translation efficiency and lower dsRNA generation compared with a conventional HEPES / Tris system, and a phosphate system can be used simultaneously with the novel All In One transcription system.

The invention relates to the technical field of biology, in particular to a promoter, a promoter flank and application of the promoter flank. Compared with the mainstream TAATACGACTCACTATAG, the promoter provided by the invention has higher yield stability and translation efficiency in different systems and coding sequences. Particularly, in a co-transcription extraction system, the yield of RNA can be obviously improved.

Disclosed are a method for designing and screening a UTR combination, a UTR combination, and a use thereof. In the method for designing and screening a UTR combination, the UTR combination comprises a 5'UTR and a 3'UTR, and the method comprises the following steps: dividing groups of the UTR combination, selecting a difference feature, and evaluating the UTR combination. The method provided by the present disclosure can simplify UTR design and screening processes, and obtain, by means of indexes and a formula, a UTR combination widely applicable to improving cell translation efficiency in organisms. A UTR combination obtained by screening according to the present invention has good translation efficiency in multiple cells. The method provided by the present invention can quickly and effectively assist in designing RNA sequences that can be efficiently translated in the field of cell therapy or mRNA vaccines.

This invention discloses a 5'UTR sequence and its application in improving mRNA translation efficiency. This invention belongs to the field of biotechnology. The 5'UTR sequence designed in this invention has excellent function in promoting mRNA expression. Compared with the commonly used HBA1 gene 5'UTR, the expression level of FLuc mRNA fused with the 5'UTR in this invention can be increased by 2 to 3.5 times.

The invention belongs to the technical field of in-vitro transcription, and particularly discloses a 5 'UTR element with a protein expression enhancing function, an expression vector and application. The 5 'UTR is selected from any one of an SPP1d element, a TSPAN1u1 element and an SPP1d-TSPAN1u1 series element; the nucleotide sequence of the SPP1d element is as shown in SEQ ID NO: 1, the nucleotide sequence of the TSPAN1u1 element is as shown in SEQ ID NO: 2, and the nucleotide sequence of the SPP1d-TSPAN1u1 tandem element is as shown in SEQ ID NO: 3. The 5 'UTR element can significantly improve translation efficiency, has a wide enhancement effect on functional protein expression, is high in biological safety, and provides a standardized and high-performance translation regulation tool for the fields of efficient recombinant protein production, gene therapy vector optimization, vaccine research and development and the like.

The invention discloses application of m6A modified circNDUFA10 in diagnosis and treatment of lung adenocarcinoma. The molecule is highly expressed in lung adenocarcinoma tissues and has a remarkable m6A modification level. The invention provides a diagnostic reagent and a kit for detecting the expression quantity and / or modification level of the gene by taking the gene as a diagnostic marker. The m6A recognition protein YTHDF3 is taken as a treatment target, antisense oligonucleotides and related drugs for inhibiting expression of the m6A recognition protein YTHDF3 are provided, and c-Myc translation efficiency is enhanced by combining with the m6A recognition protein YTHDF3, so that a c-Myc signal channel is regulated and controlled, proliferation and metastasis of lung adenocarcinoma are inhibited, and a new means is provided for accurate diagnosis and treatment of lung adenocarcinoma.

The present disclosure relates to compositions and methods to modulate the level of expression of a protein in a deliberate manner (i.e., tunable regulation of expression) with only a minimal change to the genetic sequence of the gene of interest. The present disclosure therefore also provides compositions and methods to predictably alter protein abundance.

The invention discloses a 5 '-UTR element capable of improving the mRNA translation efficiency. The nucleotide sequence of the 5'-UTR element is 5 '-GGCCACCAACGCATTCGTTTTTTCGTT-3'. According to the 5 '-UTR element, the translation efficiency of mRNA is improved by at least 1.5 times through the new and optimized 5'-UTR element, and the 5 '-UTR element can be used as an element for enhancing the mRNA expression efficiency in nucleic acid treatment drugs or mRNA vaccines, so that more and better choices are provided for mRNA therapy.

The invention relates to the technical field of medicine, in particular to a porcine circovirus type 3 Cap recombinant protein and a matched detection method and application thereof.According to the porcine circovirus type 3 Cap recombinant protein, efficient and high-quality preparation is achieved through gene optimization and expression process improvement, Escherichia coli rare codons in ORF2 genes are replaced with GeneOptimizer software, an unfavorable structure rich in arginine at the N end is removed, and the recombinant protein is obtained. The mRNA stability and the translation efficiency are obviously improved; through IPTG induction condition optimization and'inclusion body treatment-Ni-NTA affinity chromatography-dialysis renaturation 'purification processes, the final protein purity exceeds 95%, a high-quality material basis is provided for subsequent detection, the matched indirect ELISA detection method has the core characteristics of'accuracy, high efficiency and stability', detection can be completed within 2-3 h, synchronous analysis of large-batch samples is supported, and the detection efficiency is greatly improved.