39 results about "Small hairpin RNA" patented technology

Methods for modulating T-type calcium channel activity without directly targeting the T-type calcium channels are provided that include modulating kelch-like protein 1 (KLHL1) levels in a subject by providing a small hairpin RNA (shRNA) that targets a KLHL1 gene, and then administering the shRNA to the subject in an amount sufficient to modulate KLHL1 gene expression. The KLHL1 level directly effects current activity in T-type calcium channels and therefore modulation of KLHL 1 gene expression indirectly modulates current activity in T-type calcium channels. The methods may be implemented with, for example, an shRNA molecule suitable for modulating a KLHL1 level in the subject which may be provided as a plasmid encoding the shRNA molecule or an adeno-associated virus vector encoding the shRNA molecule. Methods of identifying compounds that modulate current activity in T-type calcium channels by determining an effect of the compound on KLHL1 gene expression are also provided.

The present invention relates to nucleic acids designed to prevent the binding of single strand RNA in protein complexes originatig from small interfering RNA (siRNA) or small hairpin RNA (shRNA) and uses thereof.

Methods, compositions, and kits that include small hairpin RNA (shRNA) useful for inhibition of gene expression, such as viral-mediated gene expression, are described.

The present invention provides a method of reducing levels of at least one target protein in a cell. The cell is contacted with a first agent and a second agent. The first agent reduces synthesis of the target protein, e.g., by reducing levels of the mRNA of the target protein or inhibits translation of the mRNA. The second agent accelerates degradation of the target protein. The first agent may contact the cell before, after or simultaneously with the second agent. The first agent and the second agent may be in separate delivery vehicles, or in a single delivery vehicle. The first agent may be an RNAi (RNA interference) molecule, such as a small interfering RNA (siRNA), a small hairpin RNA (shRNA) or a microRNA (miRNA). The second agent may be a chimeric polypeptide containing a ubiquitin ligase polypeptide and a target protein interacting domain. The ubiquitin ligase polypeptide can be an E3 ubiquitin ligase, including, but not limited to, an SCF polypeptide, a HECT polypeptide and a UBR1 polypeptide. In one embodiment, the SCF polypeptide is an F-box polypeptide.

In one aspect, the invention is directed to a method of characterizing a mechanism of action of a combination of agents. The method comprises contacting a plurality of populations of cells with a combination of agents to be assessed, wherein each population of cells have one gene of interest targeted by a small hairpin RNA (shRNA) and wherein the gene of interest regulates cell death and a plurality of genes that regulate cell death are targeted in the plurality of populations of cells. A responsiveness of each population of cells to the combination of agents is determined, thereby obtaining an shRNA signature of the combination of agents so as to identify one or more genes that mediate a response to the combination of agents, thereby characterizing the mechanism of action of the combination of agents. In another aspect, the invention is directed to a method of determining whether a patient population treated with a first agent would benefit from a treatment using the first agent in combination with one or more additional agents. In yet another aspect, the invention is directed to method of determining whether a formulation of one or more agents maintains a mechanism of action of the one or more agents when unformulated.

The invention belongs to the field of biopharmacy and relates to a small hairpin RNA (Ribonucleic Acid) recombination oncolytic adenovirus carrying a target human kinesin (mitotic phase phosphoprotein 1) gene and a preparation method of the recombination oncolytic adenovirus. The invention provides a brand-new liver cancer drug target, namely mitotic phase phosphoprotein 1 which has a broad spectrum that the traditional gene therapy target p53 gene does not have; the invention further provides the preparation method of the small hairpin RNA recombination oncolytic adenovirus for expressing the mitotic phase phosphoprotein 1; and at the same time, the invention provides an application of the recombination oncolytic adenovirus in preparing a cancer treatment drug.

The invention belongs to the technical field of biomedicines and molecular biology and relates to siRNA (small interference Ribonucleic Acid) for targeted inhibition on expression of an esophagus cancer EGFL6 gene, and an RNA interference recombinant lentiviral vector for the targeted inhibition on the expression of the esophagus cancer EGFL6 gene. The constructed RNA interference recombinant lentiviral vector is used to inhibit the expression of the esophagus cancer EGFL6 gene and effectively inhibit the expression of the esophagus cancer EGFL6 gene. The siRNA has double strands and is markedas si EGFL6; a deactivated lentiviral vector is used to carry a DNA (Deoxyribonucleic Acid) template of a UCP2 RNA interference segment, and is transcribed into shRNA (small hairpin RNA) in vivo, sothat the problems of targeting ability, safety and expression continuity of RNA interference are perfectly solved. A target gene is integrated to a target cell genome, is expressed for long time and has a small immune response, so that the siRNA is a relatively ideal interference vector which is introduced into tumor cells and is used for silencing the EGFL6 gene in the tumor cell.

This invention provides vector systems based on the promoters of Epstein-Barr virus-encoded small RNAs that can be used to express and deliver desired RNA molecules such as small hairpin RNAs in mammalian cells. Such small hairpin RNAs are useful for RNA interference.

This invention generally relates to a composition and its production method useful for developing drugs and / or therapies for enhancing wound healing, in particular scarless wound healing. Particularly, the present invention teaches the essential processes necessary for producing and purifying embryonic stem cell (ESC)-specific RNA compositions, such as messenger RNAs (mRNA), microRNA precursors (pre-miRNA), and small hairpin RNAs (shRNA), which are useful for treating human diseases and lesions.

Methods for modulating T-type calcium channel activity without directly targeting the T-type calcium channels are provided that include modulating kelch-like protein 1 (KLHL1) levels in a subject by providing a small hairpin RNA (shRNA) that targets a KLHL1 gene, and then administering the shRNA to the subject in an amount sufficient to modulate KLHL1 gene expression. The KLHL1 level directly effects current activity in T-type calcium channels and therefore modulation of KLHL1 gene expression indirectly modulates current activity in T-type calcium channels. The methods may be implemented with, for example, an shRNA molecule suitable for modulating a KLHL1 level in the subject which may be provided as a plasmid encoding the shRNA molecule or an adeno-associated virus vector encoding the shRNA molecule. Methods of identifying compounds that modulate current activity in T-type calcium channels by determining an effect of the compound on KLHL1 gene expression are also provided.

Disclosed are improved shRNA molecules, termed “organic shRNA” (OshRNA), that incorporate certain structural features that increase the likelihood that the desired guide strand is produced while reducing accumulation of passenger strands that might contribute to off-target effects. Also provided herein are nucleic acids encoding OshRNAs, kits, cells, and transgenic animals comprising such nucleic acids, as well as methods of making and using OshRNAs and / or nucleic acids encoding OshRNAs.

A fusion molecule is provided that includes one or more inhibitory nucleic acids, a targeting polypeptide, and a nucleic acid binding moiety. The targeting polypeptide and the nucleic acid binding moiety include specific the amino acid sequences. A fusion molecule is also provided that includes one or more inhibitory nucleic acids, a targeting polypeptide, and a nucleic acid binding moiety adapted to bind a double-stranded RNA or to a small hairpin RNA. The targeting polypeptide being CCL27 or CCL11 or a fragment thereof.

This invention relates generally to the use of ITK (IL-2 inducible T-cell kinase) inhibitors for the treatment of patients who have melanoma or other solid tumors. Applicants demonstrate that ITK lentiviral small hairpin RNA knockdowns and a small molecule inhibitor can each decrease the proliferation and migration of melanoma cell lines and a small molecule inhibitor can be used to inhibit the growth of melanomas in vivo.

The dumbbell-shaped DNA minimum vector represents a genetic vector consisting only of a gene expression cassette of interest and a terminal closed loop structure. The dumbbell vector for small hairpin RNA or microRNA expression is very small in volume, which is advantageous for cell delivery and nuclear diffusion. Conventional strategies for generating dumbbell vectors expressing small RNA entail cloning of corresponding plasmid vectors, which are then used for dumbbell protection. Herein, we provide a novel cloning-free method for generating dumbbell vectors that express small RNA, which do not require any restriction endonuclease. The method comprises performing PCR amplification on a universal DNA template using a primer containing a sense or antisense strand of a sequence of interest, denaturing and refolding the amplification product to form a stem-loop structure, and covalently closing the structure using a DNA ligase to obtain a dumbbell structure.

The present invention provides a method of reducing levels of at least one target protein in a cell. The cell is contacted with a first agent and a second agent. The first agent reduces synthesis of the target protein, e.g., by reducing levels of the mRNA of the target protein or inhibits translation of the mRNA. The second agent accelerates degradation of the target protein. The first agent may contact the cell before, after or simultaneously with the second agent. The first agent and the second agent may be in separate delivery vehicles, or in a single delivery vehicle. The first agent may be an RNAi (RNA interference) molecule, such as a small interfering RNA (siRNA), a small hairpin RNA (shRNA) or a microRNA (miRNA). The second agent may be a chimeric polypeptide containing a ubiquitin ligase polypeptide and a target protein interacting domain. The ubiquitin ligase polypeptide can be an E3 ubiquitin ligase, including, but not limited to, an SCF polypeptide, a HECT polypeptide and a UBR1 polypeptide. In one embodiment, the SCF polypeptide is an F-box polypeptide.