Nucleic acid expression construct and its use as a cell proliferation marker

Simple SummaryContent extracted from patent full text and abstract with AI.

This invention provides a nucleic acid expression construct that produces a fusion protein combining a fluorescent reporter (such as EGFP) with a protein containing a wild-type destruction signal (e.g., anillin, Aurora B kinase, Plk1, PRC1). The fusion protein localizes to key subcellular sites (cell cortex, contractile ring, midbody) during specific phases of the cell cycle and is rapidly degraded at the end of mitosis. By introducing this construct into cells (via transfection or viral transduction), cell proliferation and cell cycle progression can be precisely visualized in vitro or in vivo using fluorescence microscopy. The system can be used as a cell proliferation marker in living organisms or in culture, and transgenic animals can be generated for in vivo studies.

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

• Monitoring cell proliferation in basic cell biology research to study the cell cycle and mitosis.
• Tracking proliferating cells during embryonic development, tissue regeneration, or differentiation of stem cells.
• Analyzing cancer cell proliferation, drug efficacy, and cell cycle disruption in disease models.
• Evaluating stem cell proliferation and differentiation for regenerative medicine applications.
• Generating transgenic animals to study tissue-specific or developmental cell proliferation in vivo.
• Testing the effect of specific compounds on cell cycle kinetics and proliferation in drug discovery.
• Visualizing cell division in real time in living tissues and organisms.

BenefitsContent extracted from patent full text and abstract with AI.

• Enables high-contrast, real-time visualization of cell cycle progression at single-cell resolution.
• Fusion protein specifically marks mitotic and proliferating cells while being degraded in non-dividing/postmitotic cells, reducing background signal and false positives.
• Applicable to a broad range of organisms (human, animal, fungi, plant cells).
• Versatile use with different promoters (constitutive, cell cycle-specific, tissue/cell type-specific, or inducible), allowing targeted expression in specific cell types or at certain stages.
• Can be stably or transiently expressed in cells, or introduced via viral vectors for in vitro or in vivo studies.
• Co-localizes with established proliferation markers (e.g., Ki-67, pHH3) and can improve accuracy and convenience of proliferation assays.
• Facilitates creation of advanced transgenic models for studying proliferation in normal development, disease, and tissue repair.

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Inventors & Applicants

Patent Abstract

In a first aspect, the present invention provides a nucleic acid expression construct encoding a fusion protein comprising a fluorescence reporter protein and a protein with a wild-type destruction signal, wherein the sequence encoding the fusion protein is operably linked to a non-endogenous promoter, and wherein the fusion protein localizes during cell cycle progression to subcellular structures selected from the group consisting of the cell cortex, the contractile ring, and the midbody. In a further aspect, the present invention provides a method of visualizing cell cycle progression, said method comprising the steps of (a) introducing the nucleic acid expression construct of the present invention into a cell; (b) expressing the fusion protein encoded by the nucleic acid expression construct within the cell; (c) and monitoring the expression of the fusion protein by means of fluorescence analysis. In another aspect, the present invention provides the use of the nucleic acid expression construct of the present invention as a cell proliferation marker in vivo and in vitro. In yet another aspect, the present invention provides a transgenic animal comprising the nucleic acid expression of the present invention as well as its use for analyzing cell proliferation in vivo.