A piggyBac-based TANGO GFP assay for high throughput screening of GPCR ligands in live cells.

ABSTRACT:

Background

GPCRs are considered essential for various physiological processes and have been the most productive drug targets. Therefore, development of the methods of GPCR ligands screening is a high priority for pharmaceutical industries and research institutions.

Methods

We developed a potential method (piggyBac-TANGO) based on the TANGO and PRESTO-TANGO assays. The system was optimized with a piggyBac transposon as a transgene vehicle, and eGFP was used as a reporter instead of luciferase. The assay was validated in the HEK 293T and U87-MG cell lines and antagonist activities of the compounds were assessed. The transgene copy number and long-term stability were evaluated by qPCR. Then, we performed a DRD2-targeted screening for natural products using the piggyBac-TANGO as

SUBMITTER: Li F 

PROVIDER: S-EPMC6533772 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature