This invention provides a multi-to-multi
drug screening method for CRISPRa / i GPCR libraries based on a three-dimensional spatial
hybrid screening model and its applications, belonging to the field of
biomedical technology. The method includes the following steps: Step (1): Constructing a screening
cell pool; Step (2): Classifying the drugs to be screened; Step (3): Constructing a
drug hybrid screening model; Step (4): Grouping drugs according to mutually perpendicular X, Y, and Z planes, with drugs in each plane mixed to form independent pools; Step (5):
Processing each
pool into a
cell pool; Step (6): Detecting and analyzing the changes in sgRNA abundance in each pool; Step (7): Decoding to determine the
drug-GPCR interaction pairs. This invention offers higher screening
throughput, realizes a multi-target, multi-drug screening mode, reduces costs and time, and enables large-scale research on drug-
gene interaction networks, which is of great significance for drug research and development.