Harnessing Catalytic RNA Circuits for Construction of Artificial Signaling Pathways in Mammalian Cells

Engineering of genetic networks with artificial signaling pathways (ASPs) can reprogram cellular responses and phenotypes under different circumstances for a variety of diagnostic and therapeutic purposes. However, construction of ASPs between originally independent endogenous genes in mammalian cells is highly challenging. Here we report an amplifiable RNA circuit that can theoretically build regulatory connections between any endogenous genes in mammalian cells. We harness the system of catalytic hairpin assembly with combination of controllable CRISPR‐Cas9 function to transduce the signals from distinct messenger RNA expression of trigger genes into manipulation of target genes. Through introduction of these RNA‐based genetic circuits, mammalian cells are endowed with autonomous capabilities to sense the changes of RNA expression either induced by ligand stimuli or from various cell types and control the cellular responses and fates via apoptosis‐related ASPs. Our design provides a generalized platform for construction of ASPs inside the genetic networks of mammalian cells based on differentiated RNA expression. This study describes an amplifiable RNA circuit based on the system of catalytic hairpin assembly with combination of controllable CRISPR‐Cas9 function, which can directly build regulatory connections between originally independent endogenous genes in mammalian cells. With this design, artificial signaling pathways can be introduced into mammalian cells to control cellular responses and phenotypes through differentiated RNA expression.