Synthetic Notch Receptors for Use in Customized Spatial Control of Multiple Gene Expressions and Uses Thereof (UCLA Case No. 2023-261)

Researchers at UCLA and USC have developed a modular invention that enables customized spatial control of multiple gene expressions in living cells using synthetic Notch (synNotch) receptors, with broad applications in tissue engineering, synthetic morphogenesis, and regenerative medicine.

BACKGROUND: Precise spatial control of gene expression is fundamental to the development and function of multicellular tissues, both naturally and in tissue engineering. Traditional approaches rely on endogenous cell surface receptors and native ligands, which offer limited versatility and complex downstream signaling, restricting the ability to engineer tissues with user-defined patterns or compositions. Addressing these limitations, the invention leverages synthetic biology advances to directly program spatial gene expression within cells and tissues.

INNOVATION: The Synthetic Notch Receptor System centers on synthetic Notch receptors engineered to respond to user-defined ligands presented in spatial patterns via various materials, such as hydrogels, microparticles, elastomeric stamps, and microfluidic devices. Key features include:

• Receptors contain a customizable extracellular antigen-targeting domain, a Notch regulatory region with ligand-inducible cleavage sites, and an intracellular transcriptional module that can activate specific genes on ligand recognition.
• Ligands, presented from materials or cells, trigger cleavage of the receptor, releasing the transcriptional domain to induce expression of desired genes (including differentiation and transdifferentiation factors) within engineered spatial domains.
• Methods for presenting one or more ligands in predetermined micropatterns enable simultaneous activation of multiple gene expression programs in cells expressing distinct or multiple synNotch receptors.
• The technology supports spatial control at micron-scale resolution, allowing engineered tissues to display complex gene expression and cellular arrangements.

Benefits and Applications:

• Programmable spatial gene expression: Enables multiplexed control of cell fate decisions and tissue structure by combining multiple synthetic signaling pathways in a single construct.
• Precise tissue engineering: Supports development of engineered tissues comprising distinct cell types, such as skeletal muscle and endothelial networks, arranged in user-defined spatial patterns at cellular resolution.
• Expanded biomaterial compatibility: Functionalizes common tissue engineering scaffolds and hydrogels for gene patterning, facilitating advances in organoids, lab-grown meat, and regenerative therapies.
• Orthogonal and modular design: Receptors and ligands are fully customizable, avoiding interference with native signaling pathways and allowing for complex logic in multicellular assemblies.
• Broad utility: The toolkit enables studies in developmental biology, disease modeling, synthetic morphogenesis, and potentially clinical applications in regenerative medicine and tissue replacement.

Conclusion: This invention establishes a versatile framework for modular spatial programming of gene expression via synthetic receptors, paving the way for precise engineering of multicellular tissues and complex biological structures with unprecedented control and flexibility.