Means and methods for identifying compounds altering nuclear localization of proteins

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

This patent describes a novel cellular system for detecting and quantifying the nuclear localization of specific proteins within eukaryotic cells. The invention uses engineered cells with three genetic components: a reporter gene controlled by a response element, a transcription factor that regulates the reporter gene, and a fusion protein consisting of the protein of interest linked to a peptide that interacts with the transcription factor. When the fusion protein enters the nucleus, it triggers activation or repression of the reporter gene, producing a measurable signal (such as fluorescence or luminescence). This signal can be easily detected without microscopy, enabling high-throughput screening of compounds that affect nuclear transport of proteins.

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

• High-throughput screening for drugs that alter the nuclear import or export of proteins (e.g., in cancer or viral diseases).
• Studying mechanisms of nuclear transport and localization in basic research.
• Analyzing the effects of mutations on nuclear localization signals of proteins.
• Identifying inhibitors of viral replication by screening for compounds that prevent nuclear import of key viral proteins.
• Screening for molecules that restore proper nuclear localization of tumor suppressor proteins (e.g., p53) in cancer cell lines.
• Development of diagnostic assays to monitor nuclear or cytoplasmic localization of disease-related proteins in vitro.
• Identifying drugs that can modulate the subcellular localization of transcription factors and signaling proteins.

BenefitsContent extracted from patent full text and abstract with AI.

• Enables quantitative, fast, and high-throughput assessment of nuclear localization without reliance on microscopy.
• Can be used to screen large chemical libraries for drugs affecting nuclear transport processes, accelerating drug discovery.
• Avoids problems associated with large fluorescent tags, which can impair protein function; uses smaller peptide tags instead.
• Provides both qualitative (presence/absence) and quantitative data about protein nuclear localization.
• Adaptable to a wide range of proteins (including cellular and viral proteins) and cell types (yeast to human).
• Reporter readouts (like fluorescence/luminescence) allow for easy automation and scalability in industrial screening settings.
• Potential to identify drugs for treating cancer, viral infections, and other diseases where protein mislocalization plays a role.

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Inventors & Applicants

Patent Abstract

This invention relates to a eukaryotic cell comprising: (a) a first nucleic acid comprising a reporter gene under the control of a response element; (b) a second nucleic acid comprising a gene encoding a transcription factor, said transcription factor being capable of binding to said response element, thereby controlling expression of said reporter gene; and (c) a third nucleic acid encoding a fusion protein, said fusion protein comprising: (i) a polypeptide of interest; and (ii) a peptide being capable of binding to said transcription factor, thereby controlling the activity of said transcription factor.