An mRNA drug, which reduces expression in the liver after in vivo delivery, and a preparation method thereof. The preparation method comprises: establishing double 3′ UTR in a target mRNA, and introducing a miR-122 binding site between the double 3′ UTR, so as to effectively reduce the expression of the mRNA drug, which is delivered by means of LNP, in the liver, thereby realizing a specific inhibitory effect of miR-122 while retaining an enhancement effect of miR-122 on mRNA stability and translation efficiency. The mRNA drug can effectively reduce the expression of mRNA in the liver, realizing the efficient expression of the mRNA drug in non-hepatocellular targeted cells or tissues and increasing the effective non-hepatocyte tropism of the mRNA drug.
A cationic lipid compound, a preparation method and a use thereof, and a delivery system for mRNA are provided. The cationic lipid compound has a structure represented by formula (I):
A cationic lipid compound, a preparation method and a use thereof, and a delivery system for mRNA are provided. The cationic lipid compound has a structure represented by formula (I):
A cationic lipid compound, a preparation method and a use thereof, and a delivery system for mRNA are provided. The cationic lipid compound has a structure represented by formula (I):
In formula (I): X is O or N; n is 2-4; m is 2-4; a is 0 or 1; R1 is a chain structure comprising a tertiary amine; R2 is a linear fatty acyl group or a branched chain fatty acyl group; R3 is a branched chain fatty acyl group. The cationic lipid compound is used for preparing a delivery system for mRNA.
A61K 9/1272 - Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers
C07C 67/29 - Preparation of carboxylic acid esters by modifying the hydroxylic moiety of the ester, such modification not being an introduction of an ester group by introduction of oxygen-containing functional groups
C07C 213/06 - Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton from hydroxy amines by reactions involving the etherification or esterification of hydroxy groups
C07C 219/16 - Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having at least one of the hydroxy groups esterified by an inorganic acid or a derivative thereof
C07C 229/12 - Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of acyclic carbon skeletons
C07C 269/04 - Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups the nitrogen atom not being part of nitro or nitroso groups from amines with formation of carbamate groups
C07C 271/16 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by singly-bound oxygen atoms
C07C 271/20 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by nitrogen atoms not being part of nitro or nitroso groups
C07C 329/06 - Esters of monothiocarbonic acids having sulfur atoms of thiocarbonic groups bound to acyclic carbon atoms
C07C 333/04 - Monothiocarbamic acidsDerivatives thereof having nitrogen atoms of thiocarbamic groups bound to hydrogen atoms or to acyclic carbon atoms
C12N 15/88 - Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microencapsulation, e.g. using liposome vesicle
3.
MUTANT OF RSV PRE-FUSION CONFORMATIONAL F PROTEIN AND USE THEREOF
Provided is a mutant of a respiratory syncytial virus (RSV) pre-fusion conformational F protein. The mutant comprises an amino acid mutation relative to the amino acid sequence of a wild-type RSV F protein, the amino acid mutation being amino acids at a position 392 and a position 493 or a position near the position 493 being substituted by C, and a disulfide bond being formed between a position 392C and a position 493C or a position near the position 493C after the mutation. Also provided are a related DNA coding sequence, recombinant plasmid and mRNA sequence and a corresponding RSV vaccine. By introducing a disulfide bond at a suitable position, and combining same with cavity filling, electrostatic mutation, proline mutation and the like, the expression and stability of an F protein having pre- and post-fusion conformations and a trimerized conformation are significantly improved, and the conformation of the F protein is significantly inhibited from being converted into the post-fusion conformation. The mutant is also applied to preparation of an RSV vaccine and RSV infection detection.
Provided is an interleukin 15 fusion protein, which sequentially comprises, from N-terminal to C-terminal, an IGHE signal peptide, an IL-15 receptor Rαsushi domain, a linker peptide 1, an IL-15 functional fragment, a linker peptide 2, CH2 and CH3 fragments of an Fc monomer, wherein the amino acid at position 72 in the IL-15 functional fragment is mutated from N into D. Also provided are an mRNA of the interleukin 15 fusion protein and the use thereof. By means of optimizing the protein composition of interleukin 15 fusion protein as an IL15 superagonist, higher biological activity is achieved, so that the RLI-mono-hFc can be effectively used for treating tumors.
The present invention relates to the technical field of bioinformatics. Disclosed are a tandem design method and apparatus for multi-epitope vaccines, a device, and a storage medium. The method comprises: acquiring multiple sequence alignment data and a sequence feature matrix of candidate vaccine sequences; performing calculation according to the multiple sequence alignment data and the sequence feature matrix to obtain an initial PSSM for feature encoding, so as to obtain action feature information; at the same time, performing local feature extraction on the sequence feature matrix to obtain local feature information; obtaining an enhanced PSSM on the basis of the local feature information and the action feature information; then calculating predicted lysis probabilities of sites of the candidate vaccine sequences according to the enhanced PSSM and the sequence feature matrix; and finally constructing a mixed integer linear programming problem for optimization solution to obtain multiple multi-epitope vaccines. Thus, local hidden patterns and features of adjacent amino acid residues of the candidate vaccine sequences can be extracted, and the initial PSSM is corrected to obtain a more accurate enhanced PSSM to participate in subsequent epitope tandem design, thereby improving the accuracy and reliability of the designed multi-epitope vaccines.
122 is an alkyl group the number of C atoms of which is greater than or equal to 8, and X is O or S. The cationic lipid compounds are used for preparing the mRNA delivery system (such as LNPs), and the system has the technical effects of high delivery efficiency, good spleen targeting and high biological safety.
A61K 31/7105 - Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
An mRNA drug, which reduces expression in the liver after being delivered to the body, and a preparation method therefor. The preparation method comprises: establishing double 3'UTR in a target mRNA, and introducing a miR-122 binding site between the double 3'UTR, so as to effectively reduce the expression of the mRNA drug, which is delivered by means of LNP, in the liver, thereby realizing a specific inhibitory effect of miR-122 while retaining an enhancement effect of miR-122 on mRNA stability and translation efficiency. The mRNA drug can effectively reduce the expression of mRNA in the liver, realizing the efficient expression of the mRNA drug in non-hepatocellular targeted cells or tissues and increasing the effective non-hepatocyte tropism of the mRNA drug.
The present invention belongs to the technical field of bioinformatics. Disclosed in the present invention is a neoantigen identification method for tumor-specific circular RNAs. The method comprises: for candidate circular RNAs that have been detected, constructing pseudo-reference sequences for realignment, respectively aligning sequencing data of tumor tissue samples and paracancerous tissue samples to the pseudo-reference sequences, extracting determined first candidate reads and second candidate reads after alignment, determining candidate circular RNAs supported by the first candidate reads as first circular RNAs in the tumor tissues and candidate circular RNAs supported by the second candidate reads as second circular RNAs in the paracancerous tissues, fusing the first circular RNAs and the second circular RNAs, filtering the circular RNAs, which are present in both the tumor tissues and the paracancerous tissues, to obtain tumor-specific circular RNAs, further predicting a translation ability score, and scoring and ranking neoantigens derived from the circular RNAs to obtain top-ranked neoantigens. In the present invention, sources of tumor neoantigens can be increased, the identification accuracy of circular RNAs can also be improved, and the neoantigens thereof are more immunogenic.
Disclosed in the present application are a RSV F protein mutant and the use thereof. The amino acid sequence of the mutant relative to the wild-type RSV F protein comprises at least one amino acid mutation. The amino acid mutation is: 1) a combination of at least one engineered disulfide bond mutation and at least one cavity filling mutation; or 2) a combination of at least one engineered disulfide bond mutation, at least one cavity filling mutation, and at least one electrostatic mutation, wherein the cavity filling mutation comprises the substitution of an amino acid at position 190 with F (the numbering of the position is based on a sequence as shown in SEQ ID NO: 1), and the engineered disulfide bond mutation comprises the substitution of amino acids at positions 466 and 443 with C. Compared with the wild-type RSV F protein, the mutant can significantly increase the expression level of F having a pre-fusion conformation, and has an increased stability of the pre-fusion conformation.
The present invention relates to the technical field of biological information processing; disclosed are an mRNA sequence optimization method and apparatus, a device and a storage medium. The method comprises: determining multiple candidate sequences out of multiple mRNA sequences of the same protein; predicting a secondary structure of each candidate sequence, to obtain a secondary structure expression; annotating and parsing each secondary structure expression, to obtain secondary structure annotation information of each candidate sequence; calculating the secondary structure complexity of each candidate sequence according to the secondary structure annotation information; and finally, according to the secondary structure complexity of each candidate sequence, filtering a target mRNA sequence out of the multiple candidate sequences, such that, by means of calculating the secondary structure complexity, the secondary structure of the mRNA sequence is quantified, the reliability and effectiveness of mRNA sequence optimization may be improved, and the translation expression quantity of the mRNA sequence is improved.
1233 is a chain fatty acyl group having a branched chain. The present invention is principally used to develop a cationic lipid compound, which is used for preparing an mRNA delivery system, and has the characteristics of high delivery efficiency, organ-targeted delivery and good immune activation properties.
C07C 219/16 - Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having at least one of the hydroxy groups esterified by an inorganic acid or a derivative thereof
C07C 229/12 - Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of acyclic carbon skeletons
C07C 271/20 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by nitrogen atoms not being part of nitro or nitroso groups
C07C 333/04 - Monothiocarbamic acidsDerivatives thereof having nitrogen atoms of thiocarbamic groups bound to hydrogen atoms or to acyclic carbon atoms
C07C 329/06 - Esters of monothiocarbonic acids having sulfur atoms of thiocarbonic groups bound to acyclic carbon atoms
C07C 271/16 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by singly-bound oxygen atoms
C07C 271/22 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
C07C 269/00 - Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups the nitrogen atom not being part of nitro or nitroso groups
C07C 269/04 - Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups the nitrogen atom not being part of nitro or nitroso groups from amines with formation of carbamate groups
C07C 227/00 - Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
A61K 47/18 - AminesAmidesUreasQuaternary ammonium compoundsAmino acidsOligopeptides having up to five amino acids
A61K 47/20 - Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing sulfur, e.g. dimethyl sulfoxide [DMSO], docusate, sodium lauryl sulfate or aminosulfonic acids
A61K 9/127 - Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
12.
CATIONIC LIPID COMPOUND, PREPARATION METHOD THEREFOR AND USE THEREOF, AND MRNA DELIVERY SYSTEM
1233 is a chain fatty acyl with a branched chain. Principally developed in the present invention is a cationic lipid compound having a high delivery efficiency, organ-targeted delivery and good biological safety, which compound is used for the preparation of an mRNA delivery system and a nucleic acid drug.
C07C 219/16 - Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having at least one of the hydroxy groups esterified by an inorganic acid or a derivative thereof
C07C 229/10 - Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings
C07C 213/06 - Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton from hydroxy amines by reactions involving the etherification or esterification of hydroxy groups
C07C 329/06 - Esters of monothiocarbonic acids having sulfur atoms of thiocarbonic groups bound to acyclic carbon atoms
A61K 47/18 - AminesAmidesUreasQuaternary ammonium compoundsAmino acidsOligopeptides having up to five amino acids
A61K 47/20 - Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing sulfur, e.g. dimethyl sulfoxide [DMSO], docusate, sodium lauryl sulfate or aminosulfonic acids
A61K 9/127 - Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
13.
Method and platform for detecting immunogenicity of tumor neoantigen
A method and a platform for detecting an immunogenicity of a tumor neoantigen are provided. Specifically, the detection method includes the following steps: culturing human peripheral blood monocytes ex vivo for 13 days, adding an antigenic peptide fragment of human influenza virus and stimulating and activating cytokines, antigenic peptides, and immunoadjuvants during the 13 days, and finally conducting enzyme-linked immunospot (ELISPOT) chromogenic reaction and instrument-based scanning, counting, and analysis to detect the immunogenicity of tumor neoantigen. An application of the detection method and platform in biomedicine is provided. Compared with the prior art, the detection method and platform have advantages and characteristics of a short detection period, high convenience, low consumption of experimental cells, and low detection cost. Therefore, the detection method and platform can be used for ex vivo high-throughput assay for the immunogenicity of the tumor neoantigen.
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