The technology describes a system for labeling and targeting cells with elevated calcium levels, such as cancer cells or activated neurons, using a CREB reporter system. This system includes a synthetic CRE enhancer linked to promoters that control the expression of genes encoding functional proteins like reporters, ion channels, or therapeutic agents, enabling detection and treatment of disorders involving these cells.
Elevated calcium levels in cells are a hallmark of various pathological conditions, including cancer and brain disorders. The ability to identify and manipulate cells with dysregulated calcium is crucial for both diagnostic and therapeutic purposes. Traditional approaches often rely on non-specific markers or require prior knowledge of the cellular targets, which limits their selectivity and efficacy. Genetic manipulation techniques, while powerful, lack the precision to target specific cell types in non-transgenic organisms.
The absence of a reliable system to selectively express therapeutic agents or markers in activated cells complicates efforts to manipulate these cells for research or therapeutic purposes. Furthermore, existing methods do not adequately address the need for cell type-specificity and temporal control in gene expression, making it difficult to achieve precise targeting of dysregulated cells without affecting nearby healthy cells. These limitations underscore the need for more refined tools to detect and treat cells exhibiting aberrant calcium signaling.
The technology described is a sophisticated system for labeling and targeting cells with elevated calcium levels using a CREB reporter system. This system includes an expression vector with a bidirectional synthetic CRE enhancer linked to two promoters, each controlling a different expressible gene. The CRE enhancer contains multiple CRE palindromic sequences or CRE half sites separated by spacer sequences, which are activated by phosphorylated CREB protein in cells with dysregulated calcium, such as cancer cells or activated neurons. The promoters can be minimal or cell-type specific and may include operator elements for ligand-dependent expression control. The expressible genes can encode various functional proteins, including reporters, ion channels, cytotoxic agents, enzymes, or therapeutic proteins. This system can be used for detecting activated cells and for treating disorders involving such cells, including brain disorders and cancer.
What differentiates this technology is its dual functionality and precision in targeting specific cells based on their calcium levels. The use of a bidirectional synthetic CRE enhancer allows for simultaneous control of two genes, which can be tailored for various therapeutic and diagnostic purposes. The inclusion of cell-type-specific promoters and operator elements for ligand-dependent control adds an extra layer of specificity and control, making the system highly adaptable to different cellular environments.
Additionally, the ability to encode a wide range of functional proteins, from reporters to cytotoxic agents, provides a versatile platform for both detecting and treating a variety of conditions. This makes the technology uniquely suited for applications in complex diseases such as cancer and neurological disorders, where precise targeting and treatment of dysregulated cells are crucial.
Issued patent US11,174,495