937results about "Other foreign material introduction processes" patented technology

Aspects of the disclosure relate to complexes and other aspects relate to formulations (e.g., aqueous, lyophilized forms) comprising such complexes comprising an oligonucleotide (e.g., useful for targeting DMPK) covalently linked to an antibody (e.g., anti-TfR1 antibody).

The invention belongs to the technical field of gene engineering, and discloses a CRISPR / Cas9 delivery system and application thereof. The CRISPR / Cas9 delivery system is obtained by loading the CRISPR / Cas9 system with the fluorinated polyethyleneimine, and the fluorinated polyethyleneimine can deliver the CRISPR / Cas9 carrier into cells, has excellent transmembrane delivery efficiency, and can efficiently mediate the CRISPR / Cas9 system to enter target cells. The CRISPR / Cas9 delivery system is applied to large-scale breeding of aquatic animals, the use safety is high, the biocompatibility is excellent, and the delivery system has excellent gene editing efficiency and has wide application prospects in the field of gene editing breeding.

the present invention relates to peptides, in particular cell-penetrating peptides, having a first hydrophobic domain positioned at the C-terminus of the peptide and a second hydrophobic domain positioned at the N-terminus of the peptide, and to conjugates of such cell-penetrating peptides with a therapeutic molecule. The present invention further relates to use of such peptides or conjugates in methods of treatment or as a medicament, especially in the treatment of genetic disorders and in particular muscular dystrophies such as Duchenne muscular dystrophy.

The invention discloses an LbCas12a protein mutant as well as a preparation method and application thereof, the LbCas12a protein mutant is obtained by performing K390A or K945A mutation on a wild type LbCas12a protein, and the amino acid sequence of the wild type LbCas12a protein is as shown in SEQ ID NO. 1. According to the present invention, the LbCas12a protein is subjected to directional modification, the alanine mutation occurs at the K390 / K945 site, the LbCas12a-K390A protein mutant and the LbCas12a-K945A protein mutant are prepared, the protein mutants obtained based on the method provide a series of significant advantages in function, and the solid foundation is laid for the application of the protein mutants in multiple fields. Through Michaelis-Menten kinetic analysis, it is observed that when the LbCas12a-K390A / K945A protein mutant prepared through the method and the wild type LbCas12a protein target the same dsDNA target, the catalytic efficiency of the LbCas12a-K390A / K945A protein mutant is 42.1 times that of the wild type LbCas12a, and the catalytic efficiency of the LbCas12a-K390A / K945A protein mutant is 707.9 times that of the wild type LbCas12a, and the catalytic efficiency of the LbCas12a-K390A / K945A protein mutant is 707.9 times that of the wild type LbCas12a. The results show that K390 and K945 are mutated into alanine, so that the affinity of the LbCas12a protein to a substrate can be increased to a certain extent, and the LbCas12a protein has higher trans-cleavage activity.

The present invention features the use of cavity acoustic transducers (CATs) to apply mechanical stimuli on cells. CATs utilize the generated acoustic microstreaming vortices to trap cells and apply tunable shear on them. The present invention may use such a portable, automated, and high throughput device for cell transfection.

The present invention relates to a composition for transfecting a nucleic acid molecule into a cell and its use. The present invention relates to a composition suitable for transfecting a nucleic acid molecule into a cell, preferably a eukaryotic cell, comprising (i) at least one compound of general formula (I), preferably general formula (III), or a tautomer, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or an acceptable salt thereof, and (ii) an acceptable excipient, buffer, cell culture medium, or transfection medium, wherein Y 1 , Y 2 , Y 3 , Z 1 , Z 2 , Z 3 , X1, X2, R3, P + , R and V are as defined herein. The present invention also relates to uses of said compositions and to methods for the in vitro or ex vivo transfection of living cells.

The present invention relates to the field of non-viral vectors and pharmaceutical compositions comprising polypropyleneimine and nucleic acids, and their use in human or veterinary medicine. More particularly, the present invention relates to pharmaceutical compositions comprising polypropyleneimine polymers or copolymers for the delivery or transfection of nucleic acids, such as RNA. The pharmaceutical compositions described herein are particularly useful for (nucleic acid) vaccination, nucleic acid-based protein therapy, nucleic acid-based protein replacement therapy, gene editing, base editing, cell therapy, immunotherapy, stem cell therapy, regenerative medicine, gene silencing, nucleic acid inhibition or protein inhibition.

The present disclosure provides compositions and methods for primed editing of target DNA molecules (e.g., genomes), which enable the incorporation of nucleotide changes and / or targeted mutagenesis. Nucleotide changes can include single nucleotide changes (e.g., any transition or any transversion), the insertion of one or more nucleotides, or the deletion of one or more nucleotides. More specifically, the present disclosure provides a fusion protein comprising a nucleic acid programmable DNA binding protein (napDNAbp) and a polymerase (e.g., reverse transcriptase), which is guided to a specific DNA sequence by a modified guide RNA termed PEgRNA. The PEgRNA is modified to include an extended portion (relative to the standard guide RNA) that provides a DNA synthesis template sequence. This encodes a single-stranded DNA flap that is homologous to the strand of the targeted endogenous DNA sequence to be edited, but which contains the desired one or more nucleotide changes, and which becomes incorporated into the target DNA molecule after synthesis by the polymerase (e.g., reverse transcriptase). Various methods utilizing prime editing are also disclosed herein, including, among others, treating trinucleotide repeat contraction diseases, incorporating targeted peptide tags, treating prion diseases by incorporating protective mutations, engineering genes encoding RNA for the incorporation of RNA tags to control RNA function and expression, constructing high performance gene libraries using prime editing, inserting immune epitopes onto proteins using prime editing, using prime editing to insert inducible dimerization domains onto protein targets, and delivery methods.

The present invention is directed to a method for preparing a cell penetrating peptide-cargo formulation, said method comprising providing the cell penetrating peptide in ethanol without an aprotic solvent, associating the cell penetrating peptide with a nucleic acid cargo in an association buffer comprising 30-90% sucrose (in particular 50-90% sucrose) at a ratio of 3:1 to the cell penetrating peptide in ethanol, and storing the cell penetrating peptide-cargo formulation at -70°C or below, wherein associating is performed in a volume between 0.5x and 0.1x the final volume at which the formulation is prepared for administration. The formulation may also be lyophilised. Also provided are cell penetrating peptide-cargo formulations obtainable and / or obtained by the method herein, pharmaceutical compositions comprising the formulations, and said formulations and pharmaceutical compositions for use in the transport of the cargo across a lipid membrane and delivery of the cargo into a cell or for use in raising an immune response in a subject.

A device, system and process involve conducting electroporation of microvesicles or exosomes or other target structures in a microfluidic arrangement at pressures that exceed atmospheric pressure. Single as well as multiple flow configurations can be employed. In some cases, the system and its operation are computer-controlled for partial or complete automation.

This invention provides an effective method for the delivery and stabilization of exogenous gene materials for therapeutic purposes. [Solution] A method and composition for modulating a target genome and the stable integration of a target transgene into the cellular genome are disclosed.

The present invention pertains to the technical field of biomedicine, and specifically relates to a gene delivery system and use thereof in the preparation of a tumor therapeutic drug. The present invention relates to a technical means for inducing differentiation of tumor cells into mature cells, which utilizes messenger ribonucleic acid to regulate the expression of HNF4α in tumor cells, inhibits the malignant phenotype of malignant solid tumor cells, and achieves the effect of treating malignant solid tumors, thereby being applied to a preparation method and use of a solid tumor drug.

The invention relates to lipid nanoparticles for fish in-vivo skin gene editing and application of the lipid nanoparticles, and belongs to the technical field of fish gene editing. The lipid nanoparticle for fish in-vivo skin gene editing comprises a gene editing element and a lipid layer, wherein the lipid layer wraps the surface of the gene editing element; the gene editing element is a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 (CRISPR associated protein 9) ribonucleoprotein compound or nucleic acid for coding the compound; the lipid nanoparticle carrier comprises ionizable cationic lipid, auxiliary lipid, cholesterol, pegylated lipid and permanent cationic lipid, and the lipid nanoparticle carrier comprises the following components in parts by mole: 15 parts of ionizable cationic lipid, 25-30 parts of auxiliary lipid, 30 parts of cholesterol, 3 parts of pegylated lipid and 7 parts of permanent cationic lipid. The formula is high in pertinence, high in editing efficiency, simple, convenient and safe to operate and wide in application prospect.

Described herein are methods, compositions, kits, and synthetic peptide shuttles involving transduction of protein and / or non-protein cargoes. The methods generally comprise contacting a target eukaryotic cell with a non-protein cargo and a concentration of a synthetic peptide shuttle sufficient to increase the transduction efficiency of the non-protein cargo as compared to the absence of the synthetic peptide shuttle. In embodiments, the non-protein cargo can be a drug, such as a small molecule drug, for treating a disease. In other embodiments, novel synthetic peptide shuttles active in transducing protein and / or non-protein cargoes are described, as well as the use of propidium iodide or other membrane-impermeable fluorescent DNA intercalators as surrogate cargoes for selecting multifunctional synthetic peptide shuttles active in transducing both protein and non-protein cargoes.

The invention provides a rare single cell electroporation device and a rare single cell electroporation method for intrinsic electrical property regulation and control, and relates to the technical field of cell biology. According to the rare single cell electroporation device provided by the embodiment of the invention, real-time optimization of single cell electroporation parameters can be realized, specifically, impedance data of a single cell can be acquired based on an impedance measurement area and an impedance measurement assembly before single cell electroporation; according to the method, impedance data of a single cell is acquired, inherent electrical characteristic parameters of the single cell are calculated according to the impedance data, optimal parameters are given for the single cell, electroporation is completed based on a subsequent electroporation area and an electroporation assembly, real-time optimization of electroporation parameters is realized, and lossless and high-effectiveness rare single cell electroporation is realized.

Cas-protein-ready tau biosensor cells, CRISPR / Cas synergistic activation mediator (SAM)-ready tau biosensor cells, and methods of making and using such cells to screen for genetic modifiers of tau seeding or aggregation are provided. Reagents and methods for sensitizing such cells to tau seeding activity or tau aggregation or for causing tau aggregation are also provided.

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.

To provide compositions and methods for regulating protein production in target cells. [Solution] The compositions and methods disclosed herein can improve diseases associated with protein or enzyme deficiency. The present invention provides, for example, a composition comprising (a) at least one mRNA molecule that is at least partly encoding a functionally secreted polypeptide, and (b) a transport vehicle comprising lipid nanoparticles. The present invention also provides a method for treating a subject deficient in a functional polypeptide, comprising administering a composition comprising (a) at least one mRNA that is at least partly encoding the functionally secreted polypeptide, and (b) a transport vehicle comprising lipid nanoparticles, wherein after administration of the composition, the mRNA is expressed in a target cell to produce the functionally secreted polypeptide.

To provide the use of a compound for manufacturing a reagent for efficient in vivo delivery of nucleic acid molecules, including small RNA, to a target organ and a target cell.SOLUTION: The invention provides the use of a compound derived naturally (including a traditional Chinese medicine extract) or synthetically for the manufacture of a nucleic acid delivery reagent, where the compound is represented, e.g., by the formula in the figure.SELECTED DRAWING: None

The invention relates to an amino acid derivative carrier as well as a preparation method and application thereof, and belongs to the technical field of biology, and the preparation method of the amino acid derivative carrier comprises the following steps: providing a cationic amino acid monomer and a neutral-polyanion block polymer template; a cationic amino acid monomer, a neutral-polyanion block polymer template, a cross-linking agent and a photoinitiator are mixed and react under the irradiation of ultraviolet light to obtain an amino acid derivative carrier, and the amino acid derivative carrier is cationic nanogel. The amino acid derivative carrier prepared by the preparation method disclosed by the invention can be used as a nucleic acid drug delivery carrier, and has the advantages of strong nucleic acid loading capacity, high transfection efficiency, safety and the like.

Provided herein are compositions, systems, kits, and methods for immunizing a subject against severe fever with thrombocytopenia syndrome virus (SFTS virus) using a composition comprising: i) a plurality of nanoparticles self-assembled from a plurality of fusion proteins comprising a) at least a portion of a ferritin protein, and b) at least a portion of an immunogenic protein comprising at least a portion of the SFTS virus Gn and / or Gc envelope glycoprotein; or ii) a polynucleotide encoding the fusion protein (e.g., an mRNA sequence present in a lipid nanoparticle).

The present disclosure provides a carrier peptide fragment with excellent cell membrane permeability. The carrier peptide fragment disclosed herein includes the following amino acid sequence set forth in SEQ ID No.: 1: KKRTLRKSNRKKR, and a lysine residue at a 12th position from an N-terminus of the amino acid sequence is a D-form, and the other amino acid residues are L-forms.

While immune checkpoint block (ICB) therapy has significantly improved outcomes for metastatic melanoma, most patients do not achieve long-term benefits. Previous studies have suggested that treatment failure is partly due to insufficient immune recognition of tumor antigens (TAs). Cancer vaccines may potentially offer a complementary approach to enhance anti-tumor immunity and act synergistically with ICIs. Novel tumor antigens shared by the majority of melanoma cells are described herein. Some of the tumor antigens described herein are derived from abnormally expressed non-mutant genomic sequences, such as intronic and intergenetic sequences, which are not expressed in normal tissues. Nucleic acids, compositions, cells, and vaccines derived from these tumor antigens are described. The use of tumor antigens, nucleic acids, compositions, cells, and vaccines for the treatment of melanoma is also described.

The present invention improves the efficiency of substance introduction when a particle bombardment method is used to introduce, into a plant-derived tissue, at least one substance selected from nucleic acids and proteins for modifying a trait. A method for modifying a plant trait according to the present disclosure comprises: a step for washing a plant-derived tissue with water; a step for introducing, by using a particle bombardment method, particles retaining at least one substance selected from nucleic acids and proteins into the tissue washed with water; a step for obtaining plant bodies by growing the tissue into which the particles have been introduced; and a step for selecting a plant body of which the trait has been modified from the obtained plant bodies.

The invention discloses application of a PRTFDC1 gene in promoting porcine skeletal muscle satellite cell differentiation, myotube formation and skeletal muscle development. According to the invention, overexpression and knock-down experiments of the PRTFDC1 are respectively carried out in pig MuSCs, and immunofluorescence, qRT-PCR and Western blot analysis are carried out on cells subjected to induced differentiation, so that the PRTFDC1 is effectively shown to play a positive regulation role in a pig MuSCs differentiation process, and a new marker gene which can be applied to pig molecular breeding and growth performance evaluation is provided; and a novel molecular target and a novel detection tool are provided for muscle development regulation and biological breeding.

The invention provides a method for introducing an exogenous substance into a cell, the method comprising exposing the cell to a transient pressure reduction in the presence of the exogenous substance. Apparatus for performing the method of the invention is also provided.

To provide a method for introducing a nucleic acid into a floating cell.SOLUTION: The present invention provides a method for introducing nucleic acids into non-adherent cells, comprising the step of contacting nucleic acids with non-adherent cells in solutions comprising dextran derivatives having average molecular weights greater than 70kDa.SELECTED DRAWING: None

This invention discloses a gene editing method and application for *Trichoderma reesei* based on a ribonucleoprotein complex. It aims to overcome the problems of low editing efficiency, easy generation of heterozygotes, and risk of exogenous DNA integration in *Trichoderma reesei* using plasmid-based CRISPR / Cas9 systems. This invention combines a dual-site cleavage strategy guided by double sgRNAs with direct RNP delivery. The method involves: designing two guide RNAs spaced 50-700 bp apart for the target gene, pre-assembling them with Cas9 protein in vitro to form an active RNP complex; delivering this complex along with exogenous donor DNA to *Trichoderma reesei* protoplasts. The exogenous donor DNA contains a left homologous arm, an editing sequence, and a right homologous arm, wherein the left and right homologous arms are homologous to the genomic sequences outside the dual cleavage sites on the target gene determined by the double sgRNAs, respectively; upon entering the cell, the RNP complex introduces dual-site cleavage at the target site, achieving precise editing under the guidance of the exogenous donor DNA.