75 results about "Protein translation" patented technology

Methods and compositions for modulating repeat non-ATG protein (RAN protein) translation are provided. In some aspects, the disclosure provides methods of inhibiting RAN protein translation by contacting a cell with an effective amount of an inhibitor of eIF2 phosphorylation or an inhibitor of protein kinase R (PKR). In some embodiments, methods described by the disclosure are useful for treating diseases associated with RAN protein translation, such as certain neurodegenerative diseases.

The invention belongs to the field of bioinformatics, and relates to a protein post-translational modification prediction method based on multi-modal deep learning. The method comprises the following steps: firstly, performing multi-modal feature extraction by inputting a protein sequence and three-dimensional structure data to obtain a sequence feature vector and a structure feature vector; secondly, carrying out feature fusion by adopting a cross-modal attention mechanism and a self-adaptive gating network; then, combining the fusion features with the disease type information, and performing fine adjustment on the prediction probability through a disease specific coding network; then, using a multi-task learning framework to predict the locus probabilities of various protein post-translational modification types in parallel; finally, feature importance is calculated through a gradient back propagation technology, and a comprehensive report is output in combination with variation influence analysis. According to the method, high-precision and explainable protein post-translational modification prediction with disease perception capability is realized, and an important calculation and analysis tool is provided for revealing a disease molecular mechanism and finding accurate drug targets.

The invention discloses a multispectral intelligent identification system for protein post-translational modification sites, comprising: a spectral data acquisition module for controlling different spectrum analyzers to perform spectrum scanning and analog-to-digital conversion and formatting on protein samples in parallel to obtain multiple protein multispectral original data; the intelligent analysis module is used for carrying out preprocessing and feature extraction on the multi-spectral original data of each protein to obtain a plurality of standardized spectral feature vectors; fusing each standardized spectral feature vector to obtain a multi-dimensional feature vector, inputting the multi-dimensional feature vector into a pre-trained modification site recognition deep learning model, and outputting a recognition result list, the recognition result list comprises the positions of the recognized protein post-translation modification sites in the protein sequence and modification types corresponding to the protein post-translation modification sites; and the result output module is used for visually displaying and exporting the identification result list. According to the method, the accuracy and flux of protein post-translational modification site recognition are remarkably improved.

The invention provides a method for improving the genetic stability of PolyA tail of mRNA (messenger ribonucleic acid). The method for improving the genetic stability of the PolyA tail in the preparation process of the mRNA containing the PolyA tail comprises the steps that a plasmid vector containing three-segment PolyA polynucleotides A1-G-A2-G-A3 is transferred into a host cell to be cultured, A1, A2 and A3 are each independently composed of 30-45 continuous A, and the host cell is selected from DH5a, Stbl3 and JM109; and separating and recovering a plasmid vector containing PolyA polynucleotide, carrying out enzyme digestion linearization to obtain an mRNA transcription template, and carrying out transcription to prepare mRNA. According to the method, the high genetic stability of the plasmid PolyA tail sequence can be achieved, the stability of mRNA is not affected, the mRNA protein translation efficiency is not affected, and the Oligo-dT purification process is not affected.

This invention discloses a phosphorylated diamine morpholino oligonucleotide drug that regulates PAPOLA expression and its application, belonging to the field of biomedical technology. The nucleotide sequence of the drug is shown in SEQ ID NO:1, targeting the coding region of human PAPOLA mRNA. The formulation is a lyophilized powder for injection, with excipients including mannitol and phosphate buffer. This invention also discloses the application of this drug in the preparation of a treatment for gastric cancer, specifically advanced gastric cancer with high PAPOLA expression. The drug is administered intravenously at a dose of 8-12 mg / kg every 3 days. Its mechanism of action is to block PAPOLA protein translation, inhibit the G1 / S phase transition of gastric cancer cells, and induce apoptosis. In vitro and in vivo experiments have confirmed that this drug can specifically inhibit the proliferation and growth of gastric cancer cells, providing a new treatment option for advanced gastric cancer.

Provided herein are recombinant circular RNA (circRNA) molecules comprising an internal ribosome entry site (IRES) operably linked to a protein-coding nucleic acid sequence. The IRES may be, for example, a Type I IRES, such as a viral IRES. In some embodiments, the IRES is a synthetic IRES, such as an IRES comprising an aptamer. Methods of producing a protein in vitro or in vivo using the recombinant circRNA molecules are also provided.

The present invention provides targeting polyplexes comprised of (i) nucleic acids, in particular nucleic acids encoding pharmaceutically active peptides or proteins, and (ii) targeting conjugates comprising LPEI and PEG fragments that are connected by discrete linkages formed through defined, chemoselective reactions. Thus, the LPEI fragment is bonded in a linear end- to-end fashion to a single PEG fragment. The linear conjugates are further conjugated to a targeting fragment to enable selective interaction with a particular cell type. The polyplexes selectively deliver the nucleic acids to the targeted cells resulting in high expression and efficient protein translation as well as secretion of the encoded pharmaceutically active proteins.

This invention belongs to the field of bioinformatics and relates to a method for predicting protein post-translational modifications based on multimodal deep learning. First, multimodal feature extraction is performed on the input protein sequence and 3D structural data to obtain sequence feature vectors and structural feature vectors. Second, a cross-modal attention mechanism and an adaptive gating network are used for feature fusion. Then, the fused features are combined with disease type information, and the predicted probabilities are fine-tuned using a disease-specific coding network. Next, a multi-task learning framework is used to predict the site probabilities of various protein post-translational modification types in parallel. Finally, gradient backpropagation is used to calculate feature importance, and a comprehensive report is output based on variation impact analysis. This invention achieves high-precision, interpretable, and disease-aware protein post-translational modification prediction, providing an important computational analysis tool for revealing the molecular mechanisms of diseases and discovering precise drug targets.

The present invention pertains to the field of protein engineering, molecular imaging, molecular diagnostics, and biopharmaceutics. Provided herein are six unique Proteimer protein scaffolds (TEX-S2 / S3, TEX-S4, YTHDF3, PUM, DARPin, and Aca2) that demonstrate specificity and affinity comparable to antibodies across a broad range of therapeutic targets. Methods and protocols are provided for generating non-native protein aptamers, Proteimers, which are capable of binding to a diverse set of targets, including RNA, DNA, proteins, post-translational modifications, peptides, small molecules, and prosthetic groups. This platform supports the creation of biotherapeutic aptamers from Proteimer scaffolds for the treatment of various diseases.

The invention belongs to the field of biology, and particularly relates to construction of a recombinant Seneca virus strain with Seneca virus 2B protein translation block site mutation. The 26th lysine (K) of the 2B protein is found to be a key site for inhibiting the translation of the host protein for the first time, and after the 26th K of the 2B protein is mutated into the alanine (A), the translation retardation phenomenon of the host protein is obviously weakened; secondly, the 26th K of the 2B protein is mutated into A to construct infectious clone, and the recombinant Seneca virus strain rSVA-2B K26A with 2B protein translation retardation site mutation is successfully rescued and obtained, compared with wild type SVA, after the rSVA-2B K26A infects host cells, the translation retardation phenomenon of the host cells is weakened, the expression level of host antiviral inflammatory cytokines is enhanced, and the expression level of the recombinant Seneca virus strain rSVA-2B K26A is improved. The Seneca virus vaccine strain can be used as a potential vaccine candidate strain, and a foundation is laid for developing a novel prevention and control strategy aiming at the Seneca virus.

The invention provides a porcine low-sugar FSH (follicle stimulating hormone) recombinant protein, a recombinant expression vector and a preparation method of the porcine low-sugar FSH recombinant protein. The amino acid sequence of the swine low-sugar type FSH recombinant protein is as shown in SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3 or SEQ ID NO. 4. The invention also discloses a preparation method of the swine low-sugar type FSH recombinant protein. According to the novel swine low-sugar type FSH carrier obtained through genetic engineering, N-glycosylation motif Asn-X-Thr mutation is introduced, and N-glycosylation modification after protein translation is artificially regulated, so that stable and efficient expression of the low-sugar type FSH is realized, and the biological activity of the low-sugar type FSH carrier is also greatly improved.

Methods and compositions for modulating repeat non-ATG protein (RAN protein) translation are provided. In some aspects, the disclosure relates to methods for treating a subject having a disease associated with RAN protein translation by administering the subject a modulator of eIF3 or an eIF3 subunit, or an antibody that bind to a RAN protein.

The invention discloses application of a triple-helix structural element in RNA (Ribonucleic Acid) drug design, and belongs to the field of biological medicines. According to the application, an ENE element or a mutant of the ENE element is used as a nucleic acid stabilizing element, and the interaction between 3 'UTR and poly (A) is optimized, so that the mRNA stability is enhanced, and the protein translation efficiency is further improved, and therefore, the problem of low protein expression quantity in RNA therapy is effectively solved. The mRNA sequence optimization strategy provided by the invention provides a new method for vaccine design in the field of infectious disease and tumor immunotherapy, and has a wide application prospect.

The invention relates to a chemically modified circular RNA (Ribonucleic Acid) molecule as well as a preparation method and application thereof. Specifically, the invention discloses a chemical modification method compatible with the annular mRNA, and the immunogenicity of the annular mRNA can be remarkably reduced and the stability and the protein translation level of the annular mRNA can be remarkably enhanced by containing the chemical modification. The circular mRNA containing chemical modification can significantly improve the patent medicine property of circular RNA and reduce toxic and side effects, has wide application in the fields of vaccines, protein delivery, cell therapy and the like, and has important significance in promoting the development process of circular RNA vaccines and drugs.

The application discloses a protein post-translational modification prediction method based on multi-head attention, and belongs to the technical field of biological information, and comprises the following steps: preprocessing protein sequences with post-translational modification annotation information to obtain a legal character set, a second protein sequence data set and a benchmark data set; obtaining three kinds of vocabulary tables based on the legal character set; performing multi-scale division on protein sequence samples in the second protein sequence data set to obtain a single amino acid scale data set, a dipeptide scale data set and a tripeptide scale data set; training a corpus by using a Skip-gram model structure to obtain three kinds of word vector models; performing label prediction on protein sequence fragments of the benchmark data set by using a multi-head attention model to obtain protein sequence fragment prediction results; and the multi-scale word vector is used to comprehensively utilize polypeptide composition information in the original protein sequence, and the dependence on biological feature engineering is reduced.

A conjugate of a protein translation inhibitor, and its use in the preparation of a medicament for the treatment and / or prevention of a disease, wherein the disease is a tumor or cancer.

This invention discloses a multispectral intelligent identification system for protein post-translational modification sites, comprising: a spectral data acquisition module, which controls different spectral analysis instruments to perform parallel spectral scanning of protein samples and performs analog-to-digital conversion and formatting to obtain various raw multispectral data of proteins; an intelligent analysis module, which preprocesses and extracts features from each type of raw multispectral data of proteins to obtain various standardized spectral feature vectors; fuses each standardized spectral feature vector to obtain a multidimensional feature vector and inputs it into a pre-trained deep learning model for modification site identification, outputting a list of identification results, which includes the location of the identified protein post-translational modification sites in the protein sequence and the modification type corresponding to each protein post-translational modification site; and a result output module, which visualizes and exports the list of identification results. This invention significantly improves the accuracy and throughput of protein post-translational modification site identification.

The present invention pertains to the field of protein engineering, molecular imaging, molecular diagnostics, and biopharmaceutics. Provided herein are six unique Proteimer protein scaffolds (TEX-S2 / S3, TEX-S4, YTHDF3, PUM, DARPin, and Aca2) that demonstrate specificity and affinity comparable to antibodies across a broad range of therapeutic targets. Methods and protocols are provided for generating non-native protein aptamers, Proteimers, which are capable of binding to a diverse set of targets, including RNA, DNA, proteins, post-translational modifications, peptides, small molecules, and prosthetic groups. This platform supports the creation of biotherapeutic aptamers from Proteimer scaffolds for the treatment of various diseases.

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 relates to a plasmid vector for detecting the translation condition of amino acid sequence protein and application of the plasmid vector, and belongs to the technical field of biology. The plasmid vector for detecting the protein translation condition of the amino acid sequence, provided by the invention, is loaded with an exogenous amino acid sequence, and the amino acid sequence contains an amino acid repetitive sequence; a fluorescent protein coding gene of one color is inserted in the upstream of an amino acid repetitive sequence coding region, and a fluorescent protein coding gene of another color is inserted in the downstream of the amino acid repetitive sequence coding region. According to the invention, the translation process of the protein containing the amino acid repetitive sequence in the cell can be monitored in real time according to the expression condition of the fluorescent protein, and the formation process of the truncated object after translation blocking is captured in real time, so that a favorable tool is provided for researching and evaluating the translation condition of the protein containing the amino acid repetitive sequence in the cell; the method can also be further applied to pathogenesis research of related diseases such as Huntington's disease and / or neuronal intracellular inclusion body disease.

The invention discloses a method for constructing a vascularized tumor microenvironment based on a genetic code recoding technology, which comprises the following steps of: constructing biological orthogonal translation systems PiggyBac-(7sk-tRNAPyl) x4-EF1a-MmPylRS and PiggyBac-ETV2122 *, transfecting the biological orthogonal translation systems and transposase plasmids into a liver cancer cell line, and screening a Huh-7 cell line which stably expresses the biological orthogonal translation systems; and carrying out balling culture on the obtained Huh-7 cell line, and adding NAEK in the culture process to induce partial Huh-7 cells to be transdifferentiated into endothelial cells, so as to obtain vascularized liver cancer cell balls and realize the construction of a tumor microenvironment. The genetic code recoding technology is used for regulating and controlling formation of the functional vascular tumor microenvironment, and the method has the advantages of being simple and easy to operate and high in repeatability and universality. The technology is a near-physiological protein translation regulation and control means, the regulation and control speed is high, protein translation regulation and control can be rapidly started or stopped, a traditional two-dimensional culture mode can be replaced, multi-cell interaction is achieved, and the biomimetic property is better.

The present disclosure relates to treating mitochondrial diseases, cancer and other conditions as a result of reduced oxidative phosphorylation (OXPHOS) activity by overexpressing the METTL17 gene, encoding methyltransferase-like 17. Currently, overexpression of METTL17 to increase its copy number and / or intra-mitochondrial activity has not been indicated as a possible therapeutic for treating mitochondrial disease or other diseases such as cancer or aging related to a decline in OXPHOS activity. A variety of gene therapy approaches are presented for overexpression of METTL17 including, but not limited to, AAV, adenovirus and lentiviral vector expression.

The invention provides application of detection of enterococcus faecalis in liver cancer tissue in treatment of liver cancer. The invention finds that enterococcus faecalis is highly abundant in liver tumor tissues and is positively correlated with the pathogenesis of HCC (Hepatocellular Carcinoma). Enterococcus faecalis or an enterococcus faecalis conditioned medium promotes liver cancer cell proliferation, protein translation, cell migration and tumorigenesis. Animal experiments prove that colonization of enterococcus faecalis can promote in-vivo growth of liver cancer, and everolimus (an mTOR pathway inhibitor) administration can significantly inhibit the promotion effect. The colonization amount of the enterococcus faecalis can be used as a marker of a liver cancer clinical treatment strategy of colonization of the enterococcus faecalis.

The invention relates to the technical field of gene engineering technology, in particular to a preparation method of an efficient and stable gene knockout cell line. According to the AKAP11 gene knockout cell line provided by the invention, the 6th exon of the AKAP11 gene in the cell line is subjected to 3395bp frameshift deletion mutation, so that a coding frame is shifted, and protein translation is terminated in advance; after continuous subculture, the homozygous knockout phenotype is still maintained through PCR sequencing verification. A CRISPR / Cas9 system is combined with puromycin screening, the positive cloning rate reaches 30% or above, the construction period is shortened to 45 days, the constructed cell line can be directly used for AKAP11 related cell signal path research, disease model construction and targeted drug screening, and a reliable tool is provided for basic research and clinical transformation.

The present invention relates to DNA constructs comprising a novel TIS sequence. The TIS sequence transcribes into an RNA motif that functions as the protein translation initiation site in an mRNA transcript. The DNA construct may further comprise a nucleic acid sequence encoding signal peptide. Additionally, the DNA constructs may also comprise a nucleic acid sequence encoding a recombinant protein or one or more polypeptide chains thereof. The invention further relates to an expression vector, expression cassette and host cell which comprise said DNA constructs. Furthermore, the present invention relates to a recombinant protein expressed by the host cell as well as a method for expressing the recombinant protein.

The present application relates to the field of genetic engineering and animal genetics and breeding, in particular to a sgRNA for efficient cytosine base editing (CBE) of sheep SOCS2 gene and application thereof. The efficiency of cytosine base editing of SOCS2 gene of sheep fibroblast cells by sgRNA-S1 provided by the present application is as high as 85%; the efficiency of cytosine base editing of sheep embryos by sgRNA-S1 provided by the present application is as high as 80%, and SOCS2 gene edited sheep is obtained. The present application generates C to T mutation at the second exon of SOCS2 gene by cytosine base editing, introduces a stop codon, and prematurely terminates SOCS2 protein translation, thereby achieving efficient knock-out of SOCS2 gene and creating SOCS2 gene base edited sheep, which provides technical support for creating new germplasm of fast-growing mutton sheep and provides a new approach for creating new sheep breeds.

The invention discloses an application of protein TaeIF4A in increasing the grain number and yield of main ears of wheat. Experiments prove that the protein TaeIF4A in the overexpression A genome can significantly improve the plant height, tiller number, ear length, main ear grain number and single plant yield of wheat; according to the present invention, the coding gene TaeIF4A-6A gene of the protein TaeIF4A in the A genome and / or the coding gene TaeIF4A-6D gene of the protein TaeIF4A in the D genome are subjected to gene editing, and when the TaeIF4A-6A gene and / or the TaeIF4A-6D gene of the two homologous chromosomes are mutated to cause early termination of the protein translation, the plant height, the tiller number, the spike length, the main spike grain number and the single plant yield of the wheat can be significantly reduced; therefore, the protein TaeIF4A can regulate and control the yield, the main ear grain number, the plant height, the tiller number and / or the ear length of wheat. The method has an important application value.

The present invention provides a targeting polyplex composed of (i) a nucleic acid, particularly a nucleic acid encoding a pharmaceutically active peptide or protein, and (ii) a targeting conjugate comprising an LPEI and a PEG fragment connected by distinct linkages formed by a defined chemoselective reaction. Thus, the LPEI fragment is linked to a single PEG fragment in a linear, end-to-end manner. The linear conjugate is further conjugated to a targeting fragment to enable selective interaction with specific cell types. The polyplex selectively delivers the nucleic acid to target cells, resulting in high expression and efficient protein translation and secretion of the encoded pharmaceutically active protein.

The invention provides a method for simultaneously analyzing post-translational modifications of various proteins in a trace sample. The method comprises the following steps: acquiring a peptide fragment sample to be enriched; sequentially carrying out N-glycosylation peptide fragment enrichment, phosphorylation peptide fragment enrichment, ubiquitination peptide fragment enrichment and global peptide fragment enrichment on the to-be-enriched peptide fragment sample; wherein the peptide fragment sample to be enriched is a trace sample, and the amount of the trace sample is 1-20 [mu] g. Therefore, according to the method disclosed by the invention, streamline serial enrichment design is adopted, and enrichment steps of N-glycosylation, phosphorylation, ubiquitination modified peptide fragments and global peptide fragments are seamlessly connected, so that effluent in the previous step can be directly used for next-step enrichment, and necessary solvent evaporation and redissolution steps in a conventional process are omitted; and the loss of the trace sample in the treatment process is fundamentally reduced.

The application discloses a method for optimizing protein translation extension process to improve protein yield of saccharomyces cerevisiae, and belongs to the technical field of microbial genetic engineering. The application improves the global protein synthesis capacity of saccharomyces cerevisiae by regulating the translation elongation factor in the translation initiation process, so that the protein content of saccharomyces cerevisiae is increased from 45.63 g / 100 g to 48.06 g / 100 g, and the protein yield is increased from 3.23 g / L to 3.46 g / L. The application provides technical support for low-cost and green single-cell protein production, and has a wide application prospect in the field of protein substitution.