RT Journal Article
SR Electronic
T1 Engineering Cell Sensing and Responses Using a GPCR-Coupled CRISPR-Cas System
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 152496
DO 10.1101/152496
A1 P. C. Dave P. Dingal
A1 Nathan H. Kipniss
A1 Louai Labanieh
A1 Yuchen Gao
A1 Lei S. Qi
YR 2017
UL http://biorxiv.org/content/early/2017/06/20/152496.abstract
AB G-protein coupled receptors (GPCRs) are the largest and most diverse group of membrane receptors in eukaryotes, and detects a wide array of physiological cues in the human body. We describe a new molecular device that couples CRISPR-Cas9 programmed genome regulation to natural and synthetic extracellular signals via GPCRs. The design of our synthetic device, named CRISPR ChaCha, displays superior performance over an architecture proposed by the previously reported Tango system. Using a parsimonious mathematical model and gene-reporter assays, we find that CRISPR ChaCha can recruit and activate multiple Cas9 molecules for each GPCR molecule. We also characterize key molecular features that modulate CRISPR ChaCha performance. We adopt the design to diverse GPCRs that sense synthetic and natural ligands including chemokines, mitogens, and fatty acids, and observe efficient conversion of signals to customizable genetic programs in mammalian cells, including regulation of endogenous genes. The new class of CRISPR-coupled GPCRs provides a robust and efficient platform for engineering cells with novel behaviors in response to the diverse GPCR ligand repertoire.