Activity-Based DNA-Encoded Library Screening for Selective Inhibitors of Eukaryotic Translation.

ABSTRACT: Small molecule probes exist for only ∼2% of human proteins because most lack functional binding pockets or cannot be assayed for high-throughput screening. Selective translation modulation circumvents canonical druggability and assay development constraints by using in vitro transcription-translation (IVTT) as a universal biochemical screening assay. We developed an IVTT activity assay by fusing a GFP reporter to various target gene sequences and screened the target sequences for inhibitors in microfluidic picoliter-scale droplets using a 5,348-member translation inhibitor DNA-encoded library (DEL). Screening a proof-of-concept PCSK9-GFP reporter yielded many hits; 6/7 hits inhibited PCSK9-GFP IVTT (IC₅₀ 1-20 μM), and the lead hit reduced PCSK9 levels in HepG2 cells. Preliminary

SUBMITTER: Barhoosh H 

PROVIDER: S-EPMC11503492 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature