133results about How to "Efficient transfection" patented technology

A method is disclosed for delivering molecules such as pharmaceutical drugs and nucleic acids into skeletal muscle in vivo. The pharmaceutical drug or nucleic acid is first injected into the muscle at one or multiple sites. Immediately, or shortly after, injection, electrodes are placed flanking the injection site and a specific amount of electrical current is passed through the muscle. The electrical current makes the muscle permeable, thus allowing the pharmaceutical drug or nucleic acid to enter the cell. The efficiency of transfer permits robust immune responses using DNA vaccines and produces sufficient secreted proteins for systemic biological activity to be observed.

Disclosed are improved VP2-modified recombinant adeno-associated viral (rAAV) vectors, expression systems, and rAAV virions that are fully virulent, yet lack functional VP2 protein expression. Also disclosed are pharmaceutical compositions, virus particles, host cells, and pharmaceutical formulations that comprise these modified vectors useful in the expression of therapeutic proteins, polypeptides, peptides, antisense oligonucleotides and/or ribozymes in the cells and tissues of selected mammals, including, for example, human tissues and host cells.

Poly(amine-co-ester) polymers, methods of forming active agent-load nanoparticles therefrom, and methods of using the nanoparticles for drug delivery are disclosed. The nanoparticles can be coated with an agent that reduces surface charge, an agent that increases cell-specific targeting, or a combination thereof. Typically, the loaded nanoparticles are less toxic, more efficient at drug delivery, or a combination thereof compared to a control other transfection reagents. In some embodiments, the nanoparticles are suitable for in vivo delivery, and can be administered systemically to a subject to treat a disease or condition.

Polyamine-co-ester-co-ortho ester) polymers, methods of forming active agent-load nanoparticles therefrom, and methods of using the nanoparticles for drug delivery are disclosed. The nanoparticles can be coated with an agent that reduces surface charge, an agent that increases cell-specific targeting, or a combination thereof. Typically, the loaded nanoparticles are less toxic, more efficient at drug delivery, or a combination thereof compared to a control or other transfection reagents.

The invention discloses a recombinant adeno-associated virus-NADH dehydrogenase sigmasubunit 4 gene total length for treating Leber hereditary optic neuropathy and a medicament. The total length of the gene is shown in a nucleotide sequence SEQ ID NO:1, wherein the nucleotide sequence, consisting of a CAG starter sequence, a coded sequence of ND4 of a mitochondrial localization sequence with Cox10 and UTR with the length of 625bp, has the total length of 3824bp. The medicament is injected into an eye vitreous chamber and is used for treating Leber hereditary optic neuropathy; because the medicament can be injected into the vitreous chamber, the activity can be kept in the vitreous chamber, and the medicament can be effectively subjected to transfection to optic nerve cells; signal peptide at the front end of the protein N directionally guides the protein to enter mitochondria, and the mature protein ND4 enters the mitochondria to play a function; therefore, the Leber hereditary optic neuropathy can be effectively treated by using the medicine.

The present invention provides a controllably degradable cationic polymer for delivery of biomolecules (nucleic acids, peptides, etc.), drugs, molecules used in medical imaging applications, sensitizing agents used in cancer treatments, and molecules used in tissue engineering. The present invention also provides a method for synthesizing the polymer according to the present invention.

The invention provides the construction for a farnesyl pyrophosphoric acid synthetase RNA (Ribonucleic Acid) interference recombinant lentivirus vector, which comprises the following steps of: sieving the most effective target sequence of an FDS (farnesyl diphosphate synthase) gene RNAi (RNA interference) in a tool cell 293T cell, synthesizing the double-stranded DNA of the most effective target sequence, connecting to a pGCSIL-GFP vector and successfully constructing the recombinant vector through enzyme cutting, sequencing and identification. Researches indicate that the constructed RNA interference vector LV-sh-FDS can downwards modulate the expression of an FDS mRNA (Messenger RNA) level in a neonatal rat cardiac myocyte, simultaneously can downwards modulate the expression of myocardial hypertrophy markers such as cell areas and marker genes beta-MHC (Myosin Heavy Chain) and BNP (Brain Natriuretic Peptide), additionally can effectively inhabit the activity of RhoA while downwards modulating the FDS, can be applied in preparing medicaments for treating myocardial hypertrophy diseases and also can be applied in preparing medicaments for cholesterol metabolic control.

Provided is a method for production of nanocrystalline hydroxyapatite particles, and nanocrystalline hydroxyapatite particles produced according to the method. The nanocrystalline hydroxyapatite particles exhibit substantially superior cell transformation abilities as compared to known and commercially-available calcium phosphate kits. The nanocrystalline hydroxyapatite particles also find use in tissue engineering applications, for example bone and tooth engineering and repair applications.

Provided herein are methods and compositions related to viral vectors. Also provided herein are methods and compositions for the efficient transfection of a host, for example through the highly efficient lentivector delivery system, and for the exquisite control of the timing and level of expression of the transferred sequence of interest by the simple administration of a modulator to the host harboring the transferred sequence of interest. Also disclosed are methods of making transgenic mice and transgenic mice made using compositions and methods relating to viral vectors.

The present invention is directed compounds for reversibly modification of biologically active molecules. Described are polyconjugates systems that incorporate targeting, anti-opsonization, anti-aggregation, and transfection activities into small biocompatible in vivo delivery conjugates. The use of reversible modification provides for physiologically responsive activity modulation.

A nanostructured molecular delivery vehicle comprising magnetic materials and configured to receive passenger biomolecules. The application of a an appropriate magnetic field having a gradient orients and drives the vehicle into a biological target, which may comprise cells, cell masses, tissue slices, tissues, etc. Under the control of the magnetic field, these vehicles can penetrate cell membranes. Then, the biomolecules carried by the vehicle can be released into the cells to perform their functions. Using this “nanospearing” technique, unprecendented high transfection efficiency has been achieved in several difficult-to-transfect cells. These include, but are not limited to, Bal 17 cells, ex vivo B cells, primary cultured cortical neurons, etc. This method advances the state of the art, providing an improved technique for the introduction of exogenous molecules to cells, with the clinical applications including, but not being limited to, drug delivery, gene therapy, vaccination, etc.

Polymers including poly(amine-co-ester), poly(amine-co-amide), or a combination thereof, and nanoparticles, particularly solid core nanoparticles, formed therefrom are provided. Solid core nanoparticles fabricated from hydrophobic polymers often require the presence of cationic complexing agents to stabilize negatively charged active agents such as siRNA. However, complexing agents are optional in the disclosed formulations because the nanoparticles contain cationic amines to stabilize negatively charged nucleic acids and hydrophobic domains to condense the nucleic acid into the core of the formed nanoparticles, thus improving encapsulation efficiency. This increase in nucleic acid loading allows the disclosed solid core nanoparticles to deliver more nucleic acid per cell without increasing total polymer delivered, further reducing cytotoxicity. Pharmaceutical compositions including an effective amount of the nanoparticles are also provided, and be used, for example, for in vitro and in vivo delivery of nucleic acids.

A nucleic acid encoding a chimeric antigen receptor expressed at surface of a T lymphocyte, said chimeric antigen receptor comprises, connected in the order of, an extracellular binding domain, a transmembrane region, and an intracellular signaling domain, wherein the extracellular binding domain comprises a single chain antibody, scFv(GPC3), which specifically recognizes the C-terminal epitope of GPC3. A genetically modified T lymphocyte having a chimeric antigen receptor expressed at surface thereof, and the chimeric antigen receptor is expressed by the nucleic acid described above.

A targeted vector allowing enhanced gene transfer to human hepato-cellular carcinoma (HCC¹) cells in vitro was developed using cationic liposomes covalently conjugated with the mAb AF-20. This high affinity antibody recognizes a rapidly internalized 180 kDa cell surface glycoprotein which is abundantly expressed on the surface of human HCC and other cancer cells. Quantitative binding analysis of liposomes with target cells by flow cytometry showed specific association of mAb-targeted liposomes with human HCC cells. Using mAb-targeted cationic liposomes containing 20% DOTAP, in the presence or absence of serum, gene expression in HuH-7 cells was enhanced up to 40-fold as compared to liposomes conjugated with an isotype-matched non-relevant control antibody. Transfection specificity was not observed in a control cell line that does not express the antigen recognized by mAb AF-20. This study demonstrates that cationic liposome foormulations can be targeted with monclonal antibodies (mAbs) to enhance specific in vitro gene delivery and expression in the presence or absence of serum.

The invention discloses a positive ion nanostructure lipid carrier, a manufacturing method and the application of the positive ion nanostructure lipid carrier. A positive ion nanostructure lipid carrier compound has a structure in a general formula VI, wherein, 3A and A4 are hydrophilic head parts, 3A or A4 is a hydrophilic molecule formed by one or multiple same or different amino acids, and 3A and A4 are the same or different; R is a linking arm between the head part and a tail part and is of a chain-shaped or branched-chain structure; and 3B and B4 are hydrophobic tail parts, 3B or B4 is a chain-shaped hydrophobic molecule, and 3B and B4 are the same or different. The positive ion nanostructure lipid carrier compound disclosed by the invention has the characteristics of simple structure, convenient composition and the like; the positive ion nanostructure lipid carrier compound also has the characteristics of low toxicity and high transfection efficiency, the complicated process in composition is avoided, auxiliary lipidosome in other types is not needed to be used in a transfection process, the operation and the use are easy, thus the positive ion nanostructure lipid carrier compound disclosed by the invention has a potential development prospect of commercial transfection reagent.

The invention relates to a functionalized arborescent macro radical carrier system for targeting malignant cerebroma. The functionalized arborescent macro radical carrier system is obtained by taking one or more of the following combinations as the targeting functional factor: transferrin and transferrin monoclonal antibody, or polyclonal antibody, folic acid (FA) and epidermal growth factor receptor monoclonal antibody, or polyclonal antibody and RGD cyclic peptide, chemically connecting the combinations to arborescent macro radical polyamide-amine (PAMAM) carrier system, and then compounding the combinations with treatment factor oligonucleotide, siRNA molecule or plasmid DNA. Through intra-tumor stereotactic injection, carotid injection or tail vein, the distribution of genes in the malignant cerebroma can be improved, high expression can be realized, and the effective cerebroma inhibition can be realized. Compared with the commercialized gene carrier Oligofectamin and unmodified PAMAM, the FA-PAMAM can remarkably improve the targeting delivery ability of the in vitro and in vivo genes.

The invention discloses a method for preparing cells carrying exogenous molecules in a photoinduced perforating mode, a base material for preparing the cells and the cells, belongs to the field of medical instrument modification and the field of nanometer materials, and particularly relates to a reagent kit for modifying the surface of a cell culture base material (including a cell culture plate and other base materials capable of being used for culturing cells) with gold nano-materials (including gold nano particle aggregation, gold nano rods, gold nano cones and the like). The purpose of carrying out exogenous macromolecular transfer on cells cultured on the reagent kit is achieved through near-infrared band laser assistance. According to the system, gold nano particles are used for absorbing a large amount of laser energy at the near-infrared band so as to improve the membrane permeability of the cells cultured on the system; the activity of the cells can be well maintained by optimizing the laser power and the irradiation time; the reagent kit has the advantages of being simple, direct, fast and efficient, and can be applied to different cell lines for transfer of exogenous macromolecules of different types.