39 results about "Cytosol" patented technology

Disclosed herein is a delivery agent that facilitates effective delivery of drugs and other compounds across lipid layers. The delivery agent disclosed herein provides lipid solubility under selected conditions and aqueous solubility under other conditions, and can effectively deliver compounds into the cell cytosol. The disclosed delivery agent also avoids deleterious interactions with serum and thus provide efficient in vivo delivery of therapeutic agents.

This invention aims to create a spatially isolated local reaction microenvironment for Rubisco by introducing a complete β-carboxysome microcompartment system. This design goal avoids oxygen inhibition, laying the foundation for ultimately achieving efficient and specific CO2 fixation. It provides a novel approach to overcome the fundamental problem of low efficiency of carbon-fixing enzymes in cytosols due to intense oxygen competition. By introducing the complete carboxysome system as a module, engineered yeast strains with high carboxylase activity and significantly improved ethanol production were obtained, providing a solid foundation for building next-generation yeast cell factories that produce high-yield ethanol using CO2 as a raw material.

This invention provides targeted conjugates and methods that improve the targeting and local in vivo delivery of oligonucleotides. [Solution] A composition containing a targeted oligonucleotide-HES conjugate is provided as a method for constructing and using the conjugate in therapeutic, diagnostic, and other applications. The oligonucleotide-HES complex contained in the targeted oligonucleotide-HES conjugate can cross membranes in a receptor-independent manner and can deliver oligonucleotides containing complementary sequences to the cytosol of living cells in vivo. The targeted oligonucleotide-HES conjugate has uses including the targeting and / or local delivery of antisense oligonucleotides, siRNAs, shRNAs, Dicer substrates, miRNAs, anti-miRNAs, and other nucleic acid sequences in living organisms.

This disclosure provides an E. coli strain, which lacks thioredoxin reductase activity encoded by trxB and thioredoxin 1 activity encoded by trxA, and glutathione reductase activity encoded by gor. Said E. coli strain expresses a mutated AhpC protein having glutathione reductase activity and a cytosolic prokaryotic disulfide isomerase. The E. coli strain has an oxidative cytosol and can be used to efficiently produce proteins having disulfide bonds.

The present disclosure is directed to compositions comprising mitochondrial optogenetics-based gene therapies and their methods of use. In some embodiments, a composition described herein comprises an expression vector comprising a first nucleic acid sequence encoding a channelrhodopsin fusion protein and a second nucleic acid sequence encoding a luciferase protein. In some cases, the first nucleic acid sequence and the second nucleic acid sequence are operably linked to an expression control sequence. In some instances, the channelrhodopsin fusion protein comprises a channelrhodopsin protein linked to an inner mitochondrial membrane-mitochondrial localization signal (IMM-MLS). In some implementations, when the expression vector is expressed, the luciferase protein is localized to the cytosol. In some cases, the IMM-MLS comprises a leading sequence from a mitochondrial inner membrane protein selected from ABCB10, ABCB140, Cytochrome C, and renal outer medullary potassium channel (ROMK).

The invention relates to a conjugate for transferring an effector molecule from outside a cell into said cell, the conjugate comprising at least one effector molecule to be transferred into the cell, at least one saponin of the mono-desmosidic triterpene glycoside type or the bi-desmosidic triterpene glycoside type, and at least one single-domain antibody (sdAb), covalently bound to each other, wherein the sdAb is capable of binding to a cell-surface molecule of said cell. The invention also relates to a pharmaceutical composition comprising the conjugate of the invention. Furthermore, the invention relates to a pharmaceutical composition of the invention, for use as a medicament. In addition, the invention relates to a pharmaceutical composition of the invention, for use in the treatment or the prophylaxis of any one or more of: a cancer, an auto-immune disease such as rheumatoid arthritis, an enzyme deficiency, a disease related to an enzyme deficiency, a gene defect, a disease relating to a gene defect, an infection such as a viral infection, hypercholesterolemia, primary hyperoxaluria, haemophilia A, haemophilia B, alpha-1 antitrypsin related liver disease, acute hepatic porphyria, an amyloidosis and transthyretin-mediated amyloidosis. The invention also relates to an in vitro or ex vivo method for transferring the conjugate from outside a cell to inside said cell or for transferring the effector molecule comprised by the conjugate of the invention from outside a cell to inside said cell, preferably to the cytosol of said cell.

Compositions and methods for characterization binding proteins are described that include use of a reporting construct that incorporates a bait region that interacts with the binding protein and a reporter that is susceptible to degradation in the reporter's local environment. Complex formation between the bait region and the binding protein provides protection of the susceptible reporter from degradation. Such a reporting construct can be utilized to identify cells expressing a binding protein when the susceptible reporter is susceptible to degradation in cytosol.

Compositions and methods for treating or reducing the severity or likelihood of occurrence of a parasitic worm infection in a human, an animal, or a plant are described. Compositions including isolated, native, bioactive crystals of CryH1 or CryH13 and inactivated or live recombinant bacteria expressing a CryH1 or CryH13 crystal proteins in the cytosol of the bacterium are described. Compositions suitable for pharmaceutical and agricultural applications are provided. Methods of administering a therapeutically effective amount of a composition to a human or animal suffering from a parasitic worm infection are provided. Compositions are provided for use as animal feed, feed supplement, or in waste treatment. Methods of agricultural applications for control of parasitic worm infections are provided.

The invention discloses a gram-positive bacterium extracellular electron transport chain construction method and a synthetic microbial agent. It is found that after a lipophilic material is added into an iron reduction system, the iron reduction speed and the iron reduction amount of gram-positive bacteria with weak electron transfer capacity are remarkably increased, and the iron reduction amount of the gram-positive bacteria can be consistent with the iron reduction magnitude of gram-negative strong iron reduction bacteria. The lipophilic degree and the iron reduction rate are subjected to correlation analysis and show significant positive correlation, which reveals that the lipophilic degree and the permeable membrane capacity of the material are key parameters for determining the iron reduction capacity of gram-positive bacteria. The lipophilic material not only can smoothly penetrate through the cell wall of gram-positive bacteria, but also can be enriched on a cell membrane in quantity, and gradually permeates into a cytoplasm matrix to generate electron exchange with NADH (Nicotinamide Adenine Dinucleotide Hormone) so as to construct a gram-positive bacteria extracellular electron transport chain. The invention provides a brand-new strategy for preparing the biological geological catalytic synthetic microbial inoculum for soil iron reduction.

The present disclosure relates to a genetically modified cell capable of utilizing sucrose as energy and carbon source following the expression of a single heterologous enzyme, which upon expression is translocated from the cytosol and which is capable of hydrolysing non-phosphorylated sucrose into fructose and glucose.The identification of efficient enzymes capable of hydrolysing sucrose in its non-modified form, and which on its own enable the cell to utilize sucrose as a, or as the main and / or sole, carbon and / or energy source, i.e., without the need for multi-gene sucrose utilizing systems comprising several other heterologous polypeptides, such as other enzymes and / or transporters, is highly advantageous, since it allows for cost-effective use of sucrose in large scale production processes.

The invention relates to a method for increasing plant growth by administering phytoinitiator mRNAs obtained from seedling chloroplasts. The mRNAs can either be translated directly into chloroplasts or first translated in the cytosol and subsequently imported into the chloroplasts as proteins. In both cases, the resulting proteins lead to the activation of hormonal signaling pathways, increased protein synthesis, and a significant increase in root biomass, shoot number, and hypocotyl growth. The invention further comprises formulations and application systems such as granules, sprays, or injections that enable efficient uptake and action of the mRNAs.

The application discloses a kind of lncRNA subcellular localization prediction method based on lightGBM, comprising, first, the nucleotide of the front section multiple of known lncRNA sequence is intercepted as sequence sample one;Then by based on single strand multiclass position-specific three nucleotide bias and reverse complementary kmer, sequence sample one is respectively characterized coding, the combination of two kinds of feature coding is vector;Use lightGMB as learning algorithm;Using 5-fold cross validation optimization reverse complementary kmer and LightGBM's hyperparameter;Most intercept unknown lncRNA sequence front end multiple bases as sequence sample two, and its single strand multiclass position-specific three nucleotide bias and the combination of optimized reverse complementary kmer feature coding is input in trained lightGBM, will obtain its localization subcellular type, the patent of the application can be according to long-chain non-coding RNA sequence prediction in cytoplasm, nucleus, ribosome, cytosol, exosome five subcellular positions, the application realizes simply, and prediction precision is high.

The invention relates to a method for increasing plant growth by delivering phyto-initiator mRNAs derived from chloroplasts of seedlings. The mRNAs may either be translated directly in chloroplasts or first translated in the cytosol and subsequently imported into the chloroplasts in the form of proteins. In both cases, the resulting proteins activate hormonal signalling pathways, increased protein synthesis, and bring about a significant enhancement of root biomass, shoot number and hypocotyl growth. The invention also relates to formulations and application systems such as granules, sprays or injections which enable efficient uptake and activity of the mRNAs.

The present disclosure relates to a genetically modified cell capable of utilizing sucrose as energy and carbon source following the expression of a single heterologous enzyme, which upon expression is translocated from the cytosol and which is capable of hydrolysing non-phosphorylated sucrose into fructose and glucose.The identification of efficient enzymes capable of hydrolysing sucrose in its non-modified form, and which on its own enable the cell to utilize sucrose as a, or as the main and / or sole, carbon and / or energy source, i.e., without the need for multi-gene sucrose utilizing systems comprising several other heterologous polypeptides, such as other enzymes and / or transporters, is highly advantageous, since it allows for cost-effective use of sucrose in large scale production processes.

Compounds and methods are provided for facilitating delivery of an oligonucleotide, for example, into a nucleus and / or cytosol of a cell.

The invention relates to a skin-like chip simulation platform based on a microfluidic technology. The skin-like chip simulation platform sequentially comprises a skin-like layer, a skin-like layer, a cytoplasm matrix simulation layer and an interstitial fluid circulation layer from top to bottom, a hole array is arranged on the imitated skin layer; an interstitial fluid enrichment cavity communicated with the hole array is formed in the simulated cortex, and the interstitial fluid enrichment cavity is provided with an enrichment outlet and an enrichment inlet; an interstitial fluid micro-channel is arranged in the cytoplasm matrix simulation layer, one end of the interstitial fluid micro-channel is communicated to the enrichment inlet, and the other end of the interstitial fluid micro-channel is communicated to the interstitial fluid flowing layer; an interstitial fluid storage cavity is formed in the interstitial fluid circulation layer and is provided with a storage cavity outlet and a storage cavity inlet; the device further comprises an interstitial fluid circulation channel. The skin-like chip simulation platform designed by the invention can accurately simulate the human skin environment, provides a simulation test platform for novel extraction electrode design, parameter optimization and safety evaluation of a counter ion electroosmosis technology, and remarkably reduces the preclinical research risk and cost of new technology development.

A method of identifying a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in cytosol of a yeast cell comprising: a) providing a mutated yeast cell comprising a deletion of at least one gene of the by-pass, selected from the genes encoding the enzymes pyruvate decarboxylase, acetaldehyde dehydrogenase, and acetylCoA synthetase; b) transforming said mutated yeast cell with an expression vector comprising a heterologous nucleotide sequence encoding a candidate polypeptide having potential enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA; c) testing said recombinant mutated yeast cell for its ability to grown on minimal medium containing glucose as sole carbon source, and d) identifying said candidate polypeptide as a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in the cytosol of said yeast cell when growth of said cell is observed.

The present invention provides methods for treating a neurodegenerative disease in a subject, preventing a neurodegenerative disease in a subject, increasing neurons in a region of the brain of a subject, increasing small ubiquitin-like modifier 1 (SUMO-1) in a region of the brain of a subject, reducing Sentrin-specific protease 1 (SENP-1) in a region of the brain of a subject, reducing Serine 129 phosphorylated alpha-synuclein in a region of the brain of a subject, reducing SENP-1 nuclear translocalization from the cytosol and / or reducing protein aggregates in a region of the brain of a subject. The method comprises administering to the subject an effective amount of a pharmaceutical composition comprising a specific SENP-1 inhibitor. The specific SENP-1 inhibitor may be selected from the group consisting of SUMO-2 aldehyde, momordin lc, streptonigrin, hinokiflavone, siRNA of SENP-1, and a combination thereof. Also provided is the pharmaceutical composition.

The invention relates to a pharmaceutical combination comprising: a first conjugate comprising at least one effector molecule and a single-domain antibody (sdAb) for binding to a first cell-surface molecule; and comprising a saponin, a derivative thereof, or a second conjugate comprising a binding molecule for binding to a second cell-surface molecule and the saponin and / or the derivative thereof, wherein the saponin or the derivative thereof is a monodesmosidic or bidesmosidic triterpene glycoside. The invention also relates to a composition comprising the first conjugate and the saponin (derivative) or the second conjugate comprising the saponin (derivative). In addition, the invention relates to a pharmaceutical combination or composition of the invention, for use as a medicament, and for use in the treatment or the prophylaxis of a cancer, an auto-immune disease such as rheumatoid arthritis, an enzyme deficiency, a gene defect, a disease relating to a gene defect, an amyloidosis, a disease related to an enzyme deficiency, an infection such as a viral infection, hypercholesterolemia, primary hyperoxaluria, haemophilia A, haemophilia B, alpha-1 antitrypsin related liver disease, acute hepatic porphyria, transthyretin-mediated amyloidosis. Furthermore, the invention relates to an in vitro or ex vivo method for transferring the first conjugate of the invention from outside a cell to inside said cell, preferably to the cytosol of said cell.

Gram-negative bacterial cells for expression and secretion of heterologous peptides are disclosed. The bacterial cells include a first nucleic acid encoding a secretion signal sequence, e.g., a microcin secretion signal sequence, fused to a heterologous peptide, a second nucleic acid encoding a C39 peptidase-containing ATP-binding cassette transporter (PCAT), and a third nucleic acid encoding a membrane fusion protein. The gram-negative bacterial cells secrete the heterologous peptide from the bacterial cytosol to an external environment outside of the bacterial outer membrane. Methods for preparation and delivery of peptides to external environments are also described.

Provided herein are compositions and systems for reconstitution of RNA molecules, including methods for using these molecules. For example, such molecules can be used to deliver a protein coding sequence over two or more viral vectors (such as AAVs), resulting in reconstitution of the full-length protein in a cell. Such methods can be used to deliver a protein involved in editing a nucleic acid molecule, for example to treat a genetic disease or cancer. Also provided are compositions and systems that reduce the presence of unspliced RNA end joining (REJ) RNAs (un-joined fragments), such as those present in the cytosol. In some aspects, ribosome assembly inhibition or nuclear retention of RNA is used to reduce the presence of unspliced REJ RNAs. Also provided are methods of delivering a protein coding sequence having two or more splice forms, using at least three viral vectors (such as AAVs), resulting in reconstitution of the desired full-length protein splice variants in a cell.

Strains of yeasts are provided containing the genes for the production of cannabinoids from fatty acids. The enzymes that mediate cannabinoid production are localized to the cytosol, peroxisome or different compartments within the secretory pathway (e.g., endoplasmic reticulum, Golgi, vacuole) to ensure efficient production. The engineered microorganisms produce cannabinoids in a controlled fermentation process.

Compositions and methods for treating or reducing the severity or likelihood of occurrence of a parasitic worm infection in a human, an animal, or a plant are described. Compositions including isolated, native, bioactive crystals of CryH1 or CryH13 and inactivated or live recombinant bacteria expressing a CryH1 or CryH13 crystal proteins in the cytosol of the bacterium are described. Compositions suitable for pharmaceutical and agricultural applications are provided. Methods of administering a therapeutically effective amount of a composition to a human or animal suffering from a parasitic worm infection are provided. Compositions are provided for use as animal feed, feed supplement, or in waste treatment. Methods of agricultural applications for control of parasitic worm infections are provided.

The present invention relates to: an antibody constant region in which an endosome escape motif having activity under the slightly acidic conditions of an endosome is introduced into the constant region of an antibody, thereby imparting the ability of an immunoglobulin G (IgG) antibody to actively translocate from outside a cell to the cytoplasm; an antibody comprising the same; a method of producing the same; and uses thereof. More particularly, the present invention provides a highly efficient cytosol penetrating antibody (cytotransmab) having improved endosome escape ability by introducing an endosome escape motif into a CH3 domain, which is a heavy chain constant region of an IgG antibody, and / or a CL domain, which is a light chain constant region of an IgG antibody, and relates to a pharmaceutical composition for the treatment, prevention, or diagnosis of cancer comprising the same.

The present invention provides drug modifications for improving biodistribution and / or specificity of an anticancer drug. In certain embodiments, the compound of the invention comprises a drug, a linker and a core acid. The core acid can be varied to tune the properties of the compound within the body such that the compound more selectively distributes to tumors and is, or becomes active in the cytosol.

Process for treating a vine cultivation, which involves inserting an active yeast for winemaking use with a water residue of at least 40% into a pressurization chamber, inside which a fluid is forced at a lysis pressure greater than 1000 bar with consequent breakage of the molecular structure of the yeast and obtaining an inactivated yeast or derivative thereof containing a free amino acids fraction extracted from the cytosol of the yeast. The inactivated yeast or derivative thereof is then dissolved in a solvent in order to obtain a treatment solution, which is distributed on at least one vine plant of the vine cultivation so that the vine plant absorbs the free amino acids fraction extracted from the cytosol.

The present invention relates to methods for the production of mRNA or the protein product encoded by a gene of interest in a eukaryotic host cell, wherein the transcription is effected in the cytosol. The present invention also relates to a method for the production of recombinant Modified vaccinia virus Ankara (MVA) vector or recombinant MVA virus and methods for producing pharmaceutical compositions comprising them.

RNA therapeutics have the potential to resolve a myriad of genetic diseases. Lipid nanoparticles (LNPs) are among the most successful RNA delivery systems. Expanding their use for the treatment of more genetic diseases hinges on our ability to continuously evolve the design of LNPs with high potency, cellular-specific targeting, and low side effects. Overcoming the difficulty of releasing cargo from endocytosed LNPs remains a significant hurdle. We investigated the fundamental properties of nonviral RNA nanoparticles pertaining to the activation of topological transformations of endosomal membranes and RNA translocation into the cytosol. We showed that, beyond composition, LNP fusogenicity can be prescribed by designing LNP nanostructures that lower the energetic cost of fusion and fusion-pore formation with a target membrane. The inclusion of structurally active lipids leads to enhanced LNP endosomal fusion, fast evasion of endosomal entrapment, and efficacious RNA delivery. For example, a compound of formula I:

Compounds and methods are provided for facilitating delivery of oligonucleotides, for example, to the nucleus and / or cytosol of a cell.

Process for inducing thickening of the skin of grape berries in a vine cultivation, which involves inserting an active yeast for winemaking use with a water residue of at least 40% into a pressurization chamber, inside which a fluid is forced at a lysis pressure greater than 1000 bar to subject the active yeast to said lysis pressure with consequent breakage of the molecular structure of the yeast and obtaining an inactivated yeast or derivative thereof containing a free amino acids fraction extracted from the cytosol of the same yeast. The depressurization of the pressurization chamber then takes place and the inactivated yeast or derivative thereof is extracted from the pressurization chamber itself. Furthermore, the method involves dissolving the inactivated yeast or derivative thereof in a solvent in order to obtain a treatment solution, distributing the treatment solution on at least one vine plant of the vine cultivation and allowing the vine plant to absorb the free amino acids fraction extracted from the cytosol.