Coiled Coils as Versatile Modules for Mammalian Cell Regulation

Abstract

Synthetic biology is a rapidly growing field that allows us to better understand biological processes at the molecular level, and enables therapeutic interventions and biotechnological applications. One of the most powerful tools in synthetic biology is the small, customizable, and modular protein-protein interaction domains, which is used to regulate a wide variety of processes within mammalian cells. Here we review designed coiled coil dimers that represent a set of heterodimerization domains with many advantages. These dimers have been useful for directing the localization of selected proteins within cells, enhancing chemical or light-regulated transcription, creating fast proteolysis-based responsive systems and protein secretion, genome editing, and cell-cell interaction motifs. Additionally, we will discuss how these building blocks are used in diverse applications, such as CAR T cell regulation and genome editing. Finally, we will look at the potential for future advances in synthetic biology using these building modules.

Subunits of an E3 Ligase Complex as Degrons for Efficient Degradation of Cytosolic, Nuclear, and Membrane Proteins

“…To ensure that the substrate is connected to degrons, we used designed coiled-coil (CC) heterodimers. CC heterodimers have been previously used to build complex modular 3D structures , and logic circuits , and to regulate cellular processes. − They can be designed to be highly orthogonal to other CC pairs and natural proteins and have a tunable affinity between partner peptides . We prepared genetic fusions of all degrons with a P4 CC-forming peptide and luciferase substrate with a complementary P3 peptide (Figure a).…”

“… 29 − 31 They can be designed to be highly orthogonal to other CC pairs and natural proteins and have a tunable affinity between partner peptides. 31 We prepared genetic fusions of all degrons with a P4 CC-forming peptide and luciferase substrate with a complementary P3 peptide 32 ( Figure 2 a). By increasing the amount of cotransfected CC-degron fusion protein, we were able to modulate substrate degradation with most degrons ( Figure 2 a).…”