Means and Methods for Linking Genetic Perturbations or the Expression of a Gene or Rna of Interest with Phenotypes of Cells

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

This invention describes a method and nucleic acid constructs for linking genetic changes (such as gene editing or expression of specific RNAs) in individual cells to their observable phenotypes. It enables researchers to introduce unique genetic perturbations to each cell, label these changes with DNA/RNA barcodes, and directly map which genetic changes correspond to which cellular behaviors or characteristics, all using advanced in situ sequencing and imaging. The approach works in a wide range of cell types, does not depend on the cell's transcriptional activity, and supports high-throughput, genome-scale experiments.

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

• High-throughput screening to identify genes involved in specific signaling pathways or cellular processes.
• Functional genomics research to link specific gene modifications with observed cell behaviors in disease, drug response, or development.
• Drug discovery by identifying genetic targets affecting desired phenotypes.
• Mapping genetic determinants of cell morphology, motility, or interactions.
• Studying immune cell responses by linking gene edits to cellular activation or differentiation.
• Identifying regulators of cell fate in stem cell differentiation protocols.
• Screening for genes or RNAs that enhance production of therapeutic proteins in biotechnology.

BenefitsContent extracted from patent full text and abstract with AI.

• Enables precise mapping of genetic perturbations to phenotypic changes at the single-cell level.
• Compatible with nearly any nucleated cell type, overcoming limitations of previous in situ sequencing methods.
• Supports high cell density and very large, complex libraries; suitable for genome-scale screens.
• Works even with cells having low transcriptional activity or small size, which were problematic for older technologies.
• Allows direct combination of live-cell imaging with sequencing information for comprehensive phenotyping.
• Enables multiplexed (thousands of genes or RNAs in parallel) and quantitative optical screening.
• Can be automated and is time-efficient, reducing labor and experimental inconsistency.
• The method is highly flexible – it can be used for a variety of genetic perturbations (knockout, base editing, gene expression, etc.), and with different promoters and vectors.

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Patent Abstract

The present invention relates to a method for linking the genetic perturbations of individual cells within a cell population to the phenotype of the individual cells. The present invention also relates to a method for linking the expression of a gene or RNA of interest in individual cells within a cell population to the phenotype of the individual cells. The present invention furthermore relates to a nucleic acid molecule comprising in 5'-3' direction a U6 promoter, a gRNA and a T7 promoter reverse complement.