38results about How to "Enhanced transfection" patented technology

The invention provides a pharmaceutical agent delivery composition comprising: (i) a transport polymer comprising a linear or branched peptide having from about 10 to about 300 amino acid residues, having from about 5 to 100% histidine residues, and optionally having from 0 to about 95% non-histidine amino acid residues; (ii) at least one pharmaceutical agent; and optionally (iii) one or more intracellular delivery components in association with the transport polymer. The invention also provides methods for using such composition to deliver the pharmaceutical agent to the interior of cells.

The invention provides a branched transport polymer characterized as having at least 10 amino acids and a ratio of histidine to non-histidine amino acids greater than 1.5, said branched transport polymer comprising one or more backbones, one or more terminal branches, and optionally, one or more non-terminal branches. The branched transport polymer may be associated with a pharmaceutical agent to form a pharmaceutical agent delivery composition useful for in vivo therapies based on local injection.

The present invention provides micelles having a polynucleotide encapsulated therein, the micelle comprising copolymers comprising hydrophobic moieties in a cationic complexing block. The invention further provides methods of preparing and using said micelles, and compositions thereof.

Compositions that include nucleic acid and cationic aminoglycosides and methods for their use are provided. The subject compositions are characterized by having nucleic acid complexed with a cationic aminoglycoside, where the nucleic acid is condensed. In certain embodiments, the cationic aminoglycoside is a cationic aminoglycoside antibiotic. The composition may further include one or more of: functional groups such as targeting moieties, nuclear localization or targeting peptides, endosomolytic peptides and / or one or more lipids and / or polymers, where the lipids may be provided in a manner to encapsulate the nucleic acid. The present invention also provides methods of using and preparing the nucleic acid-aminoglycoside compositions.

Nucleic acid transfection of vascular smooth muscle cells is enhanced by the application of vibrational energy to the cells. By applying vibrational energy at frequency in the range from 1 kHz to 10 MHz and at an intensity in the range from 0.01 W / cm.sup.2 to 100 W / cm.sup.2, significant enhancement of the uptake of nucleic acids into vascular smooth muscle cells can be achieved.

The invention provides a lipid of general formula (I) or (II):wherein X1, X2 and R1 to R5 are as defined herein. Such lipids are used to form complexes with a biologically-active material such as a nucleic acid, peptide or small molecule for delivering the biologically-active material to cells. The complexes may incorporate an integrin-binding peptide and, when the biologically-active material is DNA, thereby constitute a LID complex.

The invention provides a lipid of general formula (I) or (II): wherein X1, X2 and R1 to R5 are as defined herein. Such lipids are used to form complexes with a biologically-active material such as a nucleic acid, peptide or small molecule for delivering the biologically-active material to cells. The complexes may incorporate an integrin-binding peptide and, when the biologically-active material is DNA, thereby constitute a LID complex.

Methods and compositions for delivering pharmaceutical agents into cells, in particular urothelial cells of the bladder, are provided. In the methods and compositions of the invention, a solubilized cholesterol composition is used to facilitate delivery of pharmaceutical agents. Preferably, the cholesterol is solubilized by a cyclodextrin (e.g., methyl-β-cyclodextrin) and the pharmaceutical agent comprises a polynucleotide and either a cationic lipid, a cationic polymer or a dendrimer. Improved methods for transfecting polynucleotides into cells thus are also provided, using cationic lipids, cationic polymers or dendrimers and solubilized cholesterol, wherein the transfection efficiency is enhanced compared to use of cationic lipids, cationic polymers or dendrimers alone. Preferred methods of the invention involve transfecting polynucleotides into urothelial cells, preferably for therapeutic treatment of bladder cancer using, for example, a polynucleotide(s) encoding an interleukin(s), an interferon(s), a colony stimulating factor(s) and / or a tumor suppressor(s).

The invention provides a peptide having at least 3 amino acids comprising an amino acid sequence selected froma)X1SM[SEQ.ID.NO.: 1]b)LX2HK[SEQ.ID.NO.: 2]c)PSGX3ARA[SEQ.ID.NO.: 9]d)SX4RSMNF[SEQ.ID.NO.: 16]e)LX5HKSMP[SEQ.ID.NO.: 18]in which X1 is a basic amino acid residue, X2 is Q or P, X3 is A or T, X4 is an acidic amino acid residue and X5 is P or Q.The invention further provides non-viral cell-targeting vector complexes and methods associated therewith.

Processes vectors and engineered cell lines for large-scale transfection and protein production in mammalian cells, especially Chinese Hamster Ovary (CHO) cells are described in which transfection efficiencies are realized through the use of a single vector system, the use of functional oriP sequences in all plasmids, the use of codon-optimized Epstein-Barr virus nuclear antigen-1 (EBNA1) constructs the use of a fusion protein between a truncated Epstein-Barr virus nuclear antigenen-1c (EBNA1c) protein and a herpes simplex virus protein VP16, the use of a 40 kDa fully deacetylated poly(ethylenimine) as a transfection reagent, the use of co-expression of a fibroblast growth factor (FGF) and / or the use of protein kinase B to potentiate heterologous gene expression enhancement by valproic acid (VPA).

The present invention relates to protein expression systems and in particular to a mammalian protein expression system. Specifically, the present invention provides a rodent cell line with enhanced protein production capabilities.The invention also relates to eukaryotic cloning and expression vectors and related methods, and in particular to DNA vectors capable of high level expression of a protein of interest. The invention allows for long-term episomal maintenance of expression vectors in mammalian cells.

Use of a combination of DNA expression constructs for the production of a remedy for the immunization against infections with leishmaniasis, as well as a corresponding vaccine. The abstract of the disclosure is submitted herewith as required by 37 C.F.R. §1.72(b). As stated in 37 C.F.R. §1.72(b): A brief abstract of the technical disclosure in the specification must commence on a separate sheet, preferably following the claims, under the heading “Abstract of the Disclosure.” The purpose of the abstract is to enable the Patent and Trademark Office and the public generally to determine quickly from a cursory inspection the nature and gist of the technical disclosure. The abstract shall not be used for interpreting the scope of the claims. Therefore, any statements made relating to the abstract are not intended to limit the claims in any manner and should not be interpreted as limiting the claims in any manner.

The present invention provides block copolymers comprising hydrophobic moieties in a cationic complexing block, methods of preparing the same, and intermediates thereto. The compounds are useful in a variety of applications such as the assembly of micellar polyion complexes. The invention further provides methods of using said compounds and compositions thereof.

The present invention provides optimized transfection reagents comprising mixtures of cationiclipoids. In particular, the present invention provides DNA delivery vehicles based on identifying the optimal hydrophobicity of novel cationic phospholipid derivatives that, alone or in combination, form complexes with DNA (lipoplexes) and exhibit enhanced transfection activity.

The invention provides a peptide having at least 3 amino acids comprising an amino acid sequence selected froma) X1SM[SEQ.ID.NO.: 1]b) LX2HK[SEQ.ID.NO.: 2]c) PSGX3ARA[SEQ.ID.NO.: 9]d) SX4RSMNF[SEQ.ID.NO.: 16]e) LX5HKSMP[SEQ.ID.NO.: 18]in which X1 is a basic amino acid residue, X2 is Q or P, X3 is A or T, X4 is an acidic amino acid residue and X5 is P or Q.The invention further provides non-viral cell-targeting vector complexes and methods associated therewith.

The present invention relates to methods for treatment of systemic disorders using the lung as a depot organ for transgene delivery. Transfection of the pulmonary epithelium, particularly the deep alveolar cells, or pulmonary endothelial cells, is achieved via local administration of a transgene delivery vector to the lung. The transfected cells express the transgene, and the protein thereby expressed is communicated into the circulatory system. Once entering into the circulatory system, the protein is able to achieve a systemic therapeutic effect.

A method for transfecting nucleic acid to cell comprising a step for forming a nucleic acid complex by bringing a double-stranded nucleic acid molecule into contact with a nucleic acid carrier having an amino acid sequence of alternating a basic amino acid and a hydrophobic amino acid, which has a peptide chain that forms a β-sheet structure in which a side chain of a positively charged basic amino acid is disposed on one surface side and a side chain of a hydrophobic amino acid is disposed on the opposite surface side in the presence of the double-stranded nucleic acid molecule having a double helix structure, and by binding the double-stranded nucleic acid molecule and the peptide chain through either one or both of the electrostatic interaction between the side chains of the basic amino acid and phosphate groups and hydrogen bonds between the double stranded-nucleic acid molecule and the peptide chain and a nucleic acid complex used for the same are disclosed.

Disclosed are methods for cell transfection and regulating cellular behavior. More particularly, the present disclosure relates to methods of non-viral cell transfection and regulating cellular behavior using mineral coatings that allow for the enhanced transfection of cells. The mineral coatings bind polynucleotides and provide a source of calcium and phosphate ions to enhance transfection. The present disclosure also provides a high throughput platform for screening non-viral transfection of cells. The methods of the present disclosure also provide an advantageous polynucleotide delivery platform because the mineral coatings may be deposited on various medical device materials after being specifically developed using the high throughput screening platform.