43 results about "Channelrhodopsin" patented technology

Cells are rendered sensitive to stimulation by introducing into a non-photoreceptor cell nucleic acid sequences encoding at least an opsin gene product, an arrestin gene product, and the alpha subunit of the heterotrimeric G protein of the G_q family. The introduced sequences are expressed by the cell to yield at least the opsin gene product, the arrestin gene product, and the alpha subunit of the heterotrimeric G protein of the G_q family. Retinal or a derivative thereof capable of bonding with the opsin gene product to form a rhodopsin is provided to the cell. The cell is then irradiated with light having a wavelength capable of converting the rhodopsin to metarhodopsin. The conversion of rhodopsin to metarhodopsin triggers a cascade of intracellular responses within the cell resulting in an increased intracellular concentration of IP₃ and calcium ions.

Nucleic acid vectors encoding light-gated cation-selective membrane channels, in particular channelrhodopsin-2 (Chop2), converted inner retinal neurons to photosensitive cells in photoreceptor-degenerated retina in an animal model. Such treatment restored visual perception and various aspects of vision. A method of restoring light sensitivity to a retina of a subject suffering from vision loss due to photoreceptor degeneration, as in retinitis pigmentosa or macular degeneration, is provided. The method comprises delivering to the subject by intravitreal or subretinal injection, the above nucleic acid vector which comprises an open reading frame encoding a rhodopsin, to which is operatively linked a promoter and transcriptional regulatory sequences, so that the nucleic acid is expressed in inner retinal neurons. These cells, normally light-insensitive, are converted to a light-sensitive state and transmit visual information to the brain, compensating for the loss, and leading to restoration of various visual capabilities.

The invention relates to meganuclease variants which cleave a DNA target sequence from the human Rhodopsin gene (RHO), to vectors encoding such variants, to a cell, an animal or a plant modified by such vectors and to the use of these meganuclease variants and products derived therefrom for genome therapy, ex vivo (gene cell therapy) and genome engineering including therapeutic applications and cell line engineering.

The invention, in some aspects relates to compositions and methods for altering cell activity and function and the introduction and use of light-activated ion channels.

Described herein are methods and compositions for genomic editing. Clustered regularly interspaced short palindromic (CRISPR) allows for highly selective targeting and alteration of genetic loci. Here, the Inventors demonstrate CRISPR as capable of being used in living animals to prophylactically prevent a genetic disease from manifesting. Targeting and disruption of mutated rhodopsin gene prevents progression of retinitis pigmentosa in the retinal cells of a transgenic rat model. Such techniques allow for treatment methods in subjects with dominant genetic mutations, often associated with lack of a gene product, or a toxic gene product. The described technology effectively abrogates deleterious effects due to the presence of a mutated gene copy allowing the normal function of the wild-type protein to prevent cell and vision loss. The efficacy of these in vivo mechanisms are widely extensible to similar dominant negative gene mutations causing disease, or other types of genetic disease.

CsChrimson light-activated ion channel polypeptides, their encoding polynucleotides, and variants thereof are provided. Methods of introducing and using CsChrimson light activated ion channels and variants thereof for to alter cell activity and function are also provided.

Microbial type rhodopsins, such as the light-gated cation-selective membrane channel, channelrhodopsin-2 (Chop2/ChR2) or the ion pump halorhodopsin (HaloR) are expressed in retinal ganglion cells upon transduction using recombinant AAV vectors. Selective targeting of these transgenes for expression in discrete subcellular regions or sites is achieved by including a sorting motif in the vector that can target either the central area or surround (off-center) area of these cells. Nucleic acid molecules comprising nucleotide sequences encoding such rhodopsins and sorting motifs and their use in methods of differential expression of the transgene are disclosed. These compositions and methods provide significant improvements for restoring visual perception and various aspects of vision, particular in patients with retinal disease.

The invention relates to mutant channelrhodopsins having improved properties, nucleic acid constructs encoding same, expression vectors carrying the nucleic acid construct, cells comprising said nucleic acid construct or expression vector, and their respective uses.

The present invention provides a method for constructing a structural model of a complex that a G protein-coupled protein receptor forms with a ligand capable of binding the G protein-coupled receptor and a three-dimensional structural model of an activated intermediate in the structural model of the complex. The present invention also provides a method for identifying, screening for, searching for, evaluating, or designing a ligand capable of binding a GPCR by using the three-dimensional model. In one specific method by the present invention, a three-dimensional structural model of a photoactivated intermediate of rhodopsin is constructed by using a molecule modeling software and by using the three-dimensional structural coordinate of the crystal structure of rhodopsin in such a manner that amino acid residues highly conserved among GPCRs are taken into consideration. The three-dimensional stractural model of the photoactivated intermediate of rhodopsin is subsequently used to construct structural models of activated intermediates of other GPCRs. The present invention further provides a method for identifying, screening for, searching for, evaluating, or designing a ligand that binds a GPCR to act as an agonist or an antagonist. This method employs the three-dimensional structural model constructed by the above-described method.

The invention provides compositions and kits including at least one nucleic acid or polypeptide molecule encoding for a mutant ChR2 protein. Methods of the invention include administering a composition comprising a mutant ChR2 to a subject to preserve, improve, or restore phototransduction. Preferably, the compositions and methods of the invention are provided to a subject having impaired vision, thereby restoring vision to normal levels.

The invention discloses a channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line and a construction method of the channelrhodopsin-2-green fluorescence protein (GFP) gene engineered nerve stem cell line. The cell line is the recombined nerve stem cell line C 17.2 of a channelrhodopsin-2 (ChR2) which is capable of stably expressing and a green fluorescence protein (GFP). The recombined ChR2-GFP gene engineered nerve stem cell line C 17.2 is obtained by constructing recombined slow virus, transducing C 17.2 nerve stem cell line and sifting. The channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line can be used for in vivo and in vitro studies on the functional integration of a nerve neuron of a transplanted stem cell differentiation source and a nervous system of a host and provides a favorable platform for the study on the functional integration of a nerve neuron of a transplanted stem cell differentiation and a nervous system of a host. Especially with the utilization of a filter paper digestion method, the stem cell recombining method improves the rate of GFP positive cells, is beneficial to streaming Fluorescence Activated Cell Sorting (FACS) separation, and greatly improves the efficiency of an experiment.

Methods and compositions used to identify and characterize novel rhodopsin domains, which are anion-conducting channelrhodopsins. The rhodopsin domain of these anion-conducting channelrhodopsins have been cloned, optimized and expressed in mammalian systems and thus may be used in, among others, optogenetic applications and as therapeutic agents for electrically active cell mediated disorders.

The disclosure provides non-human optogenetic animal models of depression. Specifically, non-human animals each expresses a light- responsive opsin in a neuron of the animal are provided. The animal models are useful for identifying agents and targets of therapeutic strategies for treatment of depression. Examples of using the non-human animals expressing light-responsive opsin including Halorhodopsin family of light-responsive chloride pumps and Channelrhodopsin family of light-responsive cation channel proteins are described.

Methods and compositions used to identify and characterize a new class of rhodopsins derived from algae, which are highly sensitive and efficient anion-conducting channelrhodopsins. The rhodopsin domain of these anion-conducting channelrhodopsins have been cloned and expressed in mammalian systems and thus may be used in, among others, optogenetic applications and as therapeutic agents for electrically active cell mediated disorders.

Methods and agents for suppressing expression of a mutant allele of a gene and providing a replacement nucleic acid are provided. The methods of the invention provide suppression effectors such as, for example, antisense nucleic acids, ribozymes, or RNAi, that bind to the gene or its RNA. The invention further provides for the introduction of a replacement nucleic acid with modified sequences such that the replacement nucleic acid is protected from suppression by the suppression effector. The replacement nucleic acid is modified at degenerate wobble positions in the target region of the suppression effector and thereby is not suppressed by the suppression effector. In addition, by altering wobble positions, the replacement nucleic acid can still encode a wild type gene product. The invention has the advantage that the same suppression strategy could be used to suppress, in principle, many mutations in a gene. Also disclosed is a transgenic mouse that expresses human rhodopsin (modified replacement gene) and a transgenic mouse that expresses a suppression effector targeting rhodopsin. Also disclosed in intraocular administration of siRNA.