Sander Philipp N, Gillen Miller Jared T, Lairson Luke L
Department of Chemistry, The Scripps Research Institute 10550 North Torrey Pines Road La Jolla CA 92037 USA
RSC Chem Biol. 2024 Dec 17;6(2):273-280. doi: 10.1039/d4cb00137k. eCollection 2025 Feb 5.
Based on their ability to canvas vast genetic or chemical space at low cost and high speed, DNA-encoded libraries (DEL) have served to enable both genomic and small molecule discovery. Current DEL chemical library screening approaches focus primarily on target-based affinity or activity. Here we describe an approach to record the phenotype-based activity of DNA-encoded small molecules on their cognate barcode in living cells. We transfected chloroalkane-derivatized DNA barcodes carrying photoreleasable small molecules into cells. Following photorelease, bioactive compounds induced expression of a reporter gene cassette containing self-labeling HaloTag protein that becomes covalently modified by encoding barcodes. We demonstrate that we can recover activity information from cells that received active compound following immunoprecipitation-based enrichment. This generalizable approach should enable future strategies that facilitate phenotype-based screens of DNA-encoded chemical libraries in complex cellular or organism level systems.
基于其能够以低成本和高速度筛选广阔的基因或化学空间，DNA编码文库（DEL）已被用于基因组和小分子的发现。当前的DEL化学文库筛选方法主要集中在基于靶点的亲和力或活性上。在这里，我们描述了一种在活细胞中记录DNA编码小分子基于表型的活性及其同源条形码的方法。我们将携带光可释放小分子的氯代烷烃衍生化DNA条形码转染到细胞中。光释放后，生物活性化合物诱导含有自标记卤代标签蛋白的报告基因盒的表达，该蛋白会被编码条形码共价修饰。我们证明，在基于免疫沉淀的富集后，我们可以从接受活性化合物的细胞中恢复活性信息。这种可推广的方法应该能够推动未来的策略，促进在复杂细胞或生物体水平系统中对DNA编码化学文库进行基于表型的筛选。
