Polypeptides for Light Induced Ca2 Release and Constructs Containing the Same

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This invention is a genetically engineered fusion polypeptide that combines a light-sensitive channelrhodopsin protein with a calcium-sensitive protein (such as the human RyR2 receptor), connected by a specifically designed linker. When expressed in cells, this polypeptide enables precise, light-controlled release of calcium ions (Ca2+) from intracellular stores, mimicking natural cellular calcium signaling. This system can be used in various biological models (like human cell lines or C. elegans) to study and screen the effects of different compounds on calcium-sensitive processes under physiological conditions.

Use CasesContent extracted from patent full text and abstract with AI.

• High-throughput screening of drugs that modulate calcium-sensitive proteins, particularly RyR2, for potential treatment of cardiac, neurological, and muscular diseases.
• Research into the physiological role of calcium signaling in living cells, including excitable cells like neurons and cardiomyocytes.
• Toxicity testing of compounds for effects on intracellular calcium homeostasis.
• Developing disease models in cell lines and in vivo organisms (such as C. elegans) to study disorders linked to abnormal calcium release.
• Optogenetic manipulation of calcium signals in engineered tissues, organoids, or multicellular systems for research and biomedical applications.

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• Provides precise spatial and temporal control of intracellular Ca2+ release using light, enabling more physiologically relevant experiments.
• Enables scalable, reproducible, and high-throughput screening methods for candidate drugs targeting calcium-sensitive proteins, overcoming limitations of traditional, labor-intensive methods.
• Reduces the need for non-physiological chemical treatments in assays, making results more applicable to real biological scenarios.
• Improves sensitivity and specificity in assays measuring the activity of calcium-sensitive proteins.
• Offers a universal strategy that could be adapted to other Ca2+-regulated proteins beyond RyR2.

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Inventors & Applicants

Patent Abstract

The present invention relates in a first aspect to a polypeptide comprising a channelrhodopsin moiety, a linker moiety and a moiety of a Ca2+ sensitive protein. In particular, the polypeptide is a polypeptide composed of the channelrhodopsin-2 of Chlamydomonas reinhardtii, a linker and RyR2 whereby the linker is designed to establish a distance of from 100 to 200 Angstrom between the channelrhodopsin moiety and the RyR2 moiety when present in a cellular membrane of eukaryotes. In addition, nucleic acid molecules encoding the polypeptide according to the present invention as well as vector or vector systems containing said nucleic acid molecule. Further, nucleic acid constructs comprising the nucleic acid encoding the polypeptide of the channelrhodopsin as well as the linker is provided. In addition, isolated cells, cell lines or host cells containing the vector or vector system according to the present invention or expressing the polypeptide according to the present invention are provided. Further, the C. elegans strains containing the vector according to the present invention or expressing the polypeptide according to the present invention are provided. Method for determining the activity of potential principal actives on Ca2+ sensitive proteins are provided based on determining the activity of these molecules on said Ca2+ sensitive protein.