178 results about "Translational efficiency" patented technology

The invention provides a UTR (Untranslated Region) sequence for improving mRNA (Messenger Ribonucleic Acid) stability and translation efficiency and application of the UTR sequence, and particularly provides an mRNA molecule which comprises a coding region for coding polypeptide or protein and a fragment thereof, a 5 'UTR sequence positioned at the upstream of the coding region and / or a 3' UTR sequence positioned at the downstream of the coding region, the 5 'UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 1, and the 3' UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 2; and / or, the 5 'UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 3, and the 3' UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 4. According to the present invention, the mRNA stability and the translation efficiency are improved through the new and optimized UTR sequence, and the UTR sequence can be used as the element for enhancing the RNA expression efficiency in the nucleic acid treatment drug or the mRNA vaccine so as to provide more and better choices for the mRNA therapy.

The invention relates to the technical field of biological information, in particular to a translation efficiency prediction method, a screening method, a model training method and electronic equipment. The translation efficiency prediction method is realized based on a translation efficiency prediction model, and the translation efficiency prediction model comprises an embedding module, a coding module and a prediction module. The method comprises the following steps: acquiring sequence data of messenger ribonucleic acid; extracting features of the sequence data through the embedding module to obtain feature representation of messenger ribonucleic acid; the feature representation is input into the coding module, the input feature representation is subjected to convolution processing through the coding module, a convolution result is coded based on an attention mechanism, and the updated feature representation is output according to a coding result; and performing translation efficiency prediction on the updated feature representation through the prediction module to obtain a translation efficiency prediction result. According to the embodiment of the invention, the translation efficiency of the messenger ribonucleic acid can be accurately predicted with high throughput, the screening efficiency is improved, and the screening cost is reduced.

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 embodiment of the invention discloses an mRNA sequence generation method and device, a model training method and device and electronic equipment. The method comprises the steps that a target protein sequence, a target species text corresponding to the target protein sequence and target translation efficiency corresponding to the target protein sequence are obtained; determining a first target embedding corresponding to the target species text, determining a second target embedding corresponding to the target protein sequence, and determining a third target embedding corresponding to the target translation efficiency; splicing the first target embedding, the second target embedding and the third target embedding to obtain a fourth target embedding; a sequence generation model is called to map the fourth target embedding, a target mRNA sequence is generated, and the target mRNA sequence comprises a first coding region, a first untranslated region and a second untranslated region; according to the embodiment of the invention, the accurate and complete target mRNA sequence can be obtained, the application effect of the target mRNA sequence can be improved, and the method can be widely applied to scenes such as cloud technology, artificial intelligence and smart medical treatment.

The invention discloses a method and a device for generating an mRNA sequence, electronic equipment and a storage medium. The method comprises the following steps: acquiring an amino acid sequence of a given target gene; generating an initial CDS based on the amino acid sequence of the given target gene; performing first iterative optimization on the initial CDS by adopting a reward model so as to obtain an optimized CDS; obtaining an initial UTR sequence; performing second iterative optimization on the initial UTR sequence on the basis of the optimized CDS, so as to obtain an optimized UTR sequence; and splicing the optimized UTR sequence and the optimized CDS so as to obtain a final mRNA sequence corresponding to the amino acid sequence of the given target gene. According to the method, the sequence design can be integrally optimized, the interaction among the regions is fully considered, and the stability and translation efficiency of mRNA are remarkably improved, so that the protein expression level of mRNA is improved, an efficient solution is provided for research and development of mRNA drugs, and the method has a wide application prospect in the field of pharmaceuticals.

The present invention provides a site-directed mutated influenza virus gene non-coding region nucleic acid fragment and its application in improving the expression efficiency of mRNA vaccines. The provided site-directed mutated influenza virus gene non-coding region nucleic acid fragment has a sequence of SEQ ID NO: 3. The present invention also provides an mRNA fragment for translating and expressing in a host, which contains a coding frame for encoding a target protein, and the 5′UTR and / or 3′UTR of the mRNA fragment is the site-directed mutated influenza virus gene non-coding region nucleic acid fragment. In view of the problems that the existing mRNA vaccine has low translation efficiency, relatively insufficient activation of the immune system, and difficulty in producing the expected immune protection effect, the present invention provides a method for site-directed mutagenesis of a non-coding region derived from an influenza virus M gene fragment, which can be used as the 5′UTR and / or 3′UTR of the target expression sequence, and can improve the translation efficiency of the target mRNA.

The invention belongs to the field of biotechnology and medicine, and particularly relates to mRNA (messenger Ribonucleic Acid) for coding red fluorescent protein and application thereof. The mRNA for coding the red fluorescent protein comprises an ORF sequence of the red fluorescent protein, and the nucleotide sequences of the red fluorescent protein are shown as SEQ ID NO: 5-13. According to the mRNA for coding the red fluorescent protein provided by the invention, the mRNA for coding the red fluorescent protein has relatively high translation efficiency by reasonably optimizing the nucleic acid sequence of the functional region, so that the output of the red fluorescent protein is increased, and the mRNA for coding the red fluorescent protein has a wide application prospect in the field of gene drug development.

The invention discloses a key effect and a targeted intervention strategy of YTHDF1 in regulation and control of a hepatocellular carcinoma immunosuppression microenvironment. In the tumor-associated fibroblasts, YTHDF1 enhances the translation efficiency of OTUD5 mRNA through an m6A dependency mechanism, an up-regulated deubiquitination enzyme OTUD5 is directly combined with a tryptophan metabolic rate-limiting enzyme IDO1, ubiquitination degradation of the tryptophan metabolic rate-limiting enzyme IDO1 is inhibited, tryptophan is further driven to catabolism to generate kynurenine, regulatory T cell differentiation is promoted, and an immunosuppression microenvironment is formed. Spatial transcriptomics shows that YTHDF1 + CAFs and Tregs are significantly co-localized in tumor tissues and have metabolic interaction. Based on artificial intelligence virtual screening, it is found that a natural compound hesperidin can be specifically combined with a YTH structural domain of YTHDF1, an OTUD5-IDO1 signal axis mediated by the natural compound hesperidin is blocked, and the KYN level is remarkably reduced. Animal experiments prove that the tumor inhibition rate of combined treatment of hesperidin and anti-PD-1 is increased by 40% or above compared with that of a single drug group, and synergistic interaction is achieved by inhibiting Tregs infiltration and enhancing the activity of cytotoxic T lymphocytes.

The invention relates to the technical field of solid tumor immunotherapy, in particular to an IL-2mRNA multichannel delivery method for solid tumor immunotherapy, which comprises the following steps: designing a DNA template, using a plasmid containing an IL-2 gene sequence such as human IL-2cDNA, adding a T7 promoter, a 5'untranslated region UTR and a 3 'UTR at two ends to enhance translation efficiency, carrying out in-vitro transcription through T7 RNA polymerase, and carrying out in-vitro transcription through T7 RNA polymerase. And adding a 5 '-end cap analogue such as CleanCapAG to realize co-transcription cap adding, and then preparing lipid nanoparticle LNP encapsulation and carrying out surface modification. According to the IL-2mRNA multi-channel delivery method for solid tumor immunotherapy, through multi-channel delivery design, targeting optimization and combined treatment cooperation, the toxicity problem of a traditional IL-2 therapy and the drug resistance of MHC-I defective tumors are remarkably solved, meanwhile, efficient conversion is achieved through existing equipment, IL-2 expression can be accurately regulated and controlled, the immune microenvironment can be remodeled, and the immunotherapy effect is good. Meanwhile, the system is compatible with various treatment means such as chemotherapy, radiotherapy and immune checkpoint inhibitors.

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.

To provide RNA polynucleotides that improve the translation efficiency of an RNA encoding a payload, and / or the expression of a polypeptide payload encoded by the RNA, and methods of making and using the same.SOLUTION: Disclosed herein are RNA polynucleotides comprising a 5' Cap, a 5' UTR comprising a cap proximal sequence, and a sequence encoding a payload. Also disclosed herein are compositions and medical preparations comprising the same, and methods of making and using the same.SELECTED DRAWING: None

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 provides an mRNA5 'UTR optimization method and system and a storage medium, and relates to the technical field of mRNA vaccine design, and the method comprises the following steps: obtaining a plurality of 5' UTR sequences; performing feature extraction on the 5 'UTR sequence to obtain a sequence feature of the 5' UTR sequence; obtaining a tpm value of each 5 'UTR sequence according to the sequence features through a tpm prediction model; all the 5 'UTR sequences serve as a current population, the tpm values of the 5' UTR sequences serve as target function values, the current population is optimized according to the target function values through a genetic algorithm, and the optimized 5 'UTR sequences are obtained. According to the method, the expression level of the 5 'UTR sequence is improved through iterative optimization, and the translation efficiency and expression level of mRNA are accurately improved on the basis of reducing the time consumption and cost of experimental screening.

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 relates to the technical field of vaccine preparation, and discloses a method for preparing a BVDV-1 vaccine by a non-cap-dependent mRNA (messenger ribonucleic acid) technology, which is characterized in that an IRES sequence of CVB3 is inserted into the upstream of a BVDV-1E2 gene to replace a 5'end cap structure of the traditional mRNA, so that the production process is obviously simplified, the dependence on expensive enzymatic reaction is reduced, the vaccine production cost is reduced, and the production efficiency is improved. The method is especially suitable for large-scale economic sensitive scenes such as animal husbandry. According to the present invention, the non-capped mRNA is encapsulated by using the lipid nanoparticle (LNP) with the specific molar ratio, such that the stability of the mRNA and the target cell delivery efficiency are effectively improved, the immunogenicity limitation caused by insufficient translation efficiency of the non-capped mRNA is overcome, and the immune response equivalent to the commercial vaccine is achieved in the large animal. The safety advantage is particularly prominent, inflammatory response or abnormal hemogram is not observed in all animal models, a novel solution with high efficiency, safety and economical efficiency is provided for prevention and control of livestock epidemic diseases, and the method is particularly suitable for animal infectious diseases such as bovine viral diarrhea needing long-term and extensive immunization.

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 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 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.

The invention discloses an mRNA (messenger Ribonucleic Acid) vector for in-vitro transcription as well as a construction method and application thereof, and relates to the technical field of gene engineering, and the technical key point is that the mRNA vector comprises the following gene segments: 5 'UTR (Untranslated Region), signal peptide, 3' UTR and polyadenylate tail; 5 'UTR is 70 nt or Ces1d; the signal peptide is Igk or tPAP22A; the 3 'UTR is ApoA-II or AP3B1; the polyadenylic acid is PolyA (40 + 10 + 60) or PolyA (30 + 10 + 70); the invention further provides a construction method of the mRNA vector recombinant plasmid, a cell containing the mRNA vector, a recombinant bacterium containing the mRNA vector, and an application of the mRNA vector, the cell and the recombinant bacterium in in-vitro transcription. According to the mRNA vaccine vector provided by the invention, by optimizing the vector element, the stability and translation efficiency of the vector element can be improved, meanwhile, the vaccine efficacy is improved, and an important technical support is provided for future vaccine development and disease prevention and control.

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 embodiment of the invention discloses an mRNA translation efficiency prediction method and device, a model training method and device and electronic equipment, and the method comprises the steps that a target mRNA sequence and a target species text are obtained, and the target mRNA sequence comprises a first coding region, a first untranslated region and a second untranslated region; determining first target inserts corresponding to the first coding area, the first non-translated area and the second non-translated area, and splicing each first target insert with the corresponding first prompt insert to obtain a corresponding second target insert; determining a third target embedding corresponding to the target species text, splicing the third target embedding and the plurality of second target embedding to obtain a fourth target embedding, and calling a translation efficiency prediction model to generate target translation efficiency; according to the embodiment of the invention, the method can improve the accuracy of predicting the translation efficiency of mRNA, and can be widely applied to the scenes of cloud technology, artificial intelligence, smart medical treatment and the like.

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 invention belongs to the technical field of biology, and particularly relates to a trichoderma reesei expression site H2 and application thereof as well as a recombinant trichoderma reesei strain for producing thaumatin II protein as well as a construction method and application of the recombinant trichoderma reesei strain. The Trichoderma reesei expression site H2 provided by the invention is located at the 210th site in a gene segment as shown in SEQ ID NO.6 in a Trichoderma reesei genome, and an expression level equivalent to that of a cbh1 site can be obtained by inserting an exogenous gene into the site. The recombinant trichoderma reesei strain for producing the thaumatin II protein provided by the invention contains a gene for coding the thaumatin II protein, which has a nucleotide sequence shown as SEQ ID NO.1, and the gene has high translation efficiency in the trichoderma reesei, so that the recombinant trichoderma reesei strain can efficiently produce the thaumatin II protein, and the proportion of the thaumatin II protein in the obtained total protein is high; the method has important significance in reducing the production cost of the thaumatin II protein and promoting the application of the thaumatin II protein in the food industry.