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Screening of DNA-Encoded Small Molecule Libraries inside a Living Cell.
- Source :
-
Journal of the American Chemical Society [J Am Chem Soc] 2021 Feb 24; Vol. 143 (7), pp. 2751-2756. Date of Electronic Publication: 2021 Feb 12. - Publication Year :
- 2021
-
Abstract
- DNA-encoded small molecule libraries (DELs) have facilitated the discovery of novel modulators of many different therapeutic protein targets. We report the first successful screening of a multimillion membered DEL inside a living cell. We demonstrate a novel method using oocytes from the South African clawed frog Xenopus laevis . The large size of the oocytes of 1 μL, or 100 000 times bigger than a normal somatic cell, permits simple injection of DELs, thus resolving the fundamental problem of delivering DELs across cell membranes for in vivo screening. The target protein was expressed in the oocytes fused to a prey protein, to allow specific DNA labeling and hereby discriminate between DEL members binding to the target protein and the endogenous cell proteins. The 194 million member DEL was screened against three pharmaceutically relevant protein targets, p38α, ACSS2, and DOCK5. For all three targets multiple chemical clusters were identified. For p38α, validated hits with single digit nanomolar potencies were obtained. This work demonstrates a powerful new approach to DEL screening, which eliminates the need for highly purified active target protein and which performs the screening under physiological relevant conditions and thus is poised to increase the DEL amenable target space and reduce the attrition rates.
- Subjects :
- Acetate-CoA Ligase chemistry
Acetate-CoA Ligase genetics
Acetate-CoA Ligase metabolism
Animals
Humans
Mitogen-Activated Protein Kinase 14 chemistry
Mitogen-Activated Protein Kinase 14 genetics
Mitogen-Activated Protein Kinase 14 metabolism
Oocytes metabolism
Small Molecule Libraries chemistry
Xenopus laevis growth & development
DNA metabolism
Small Molecule Libraries metabolism
Xenopus laevis metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1520-5126
- Volume :
- 143
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Journal of the American Chemical Society
- Publication Type :
- Academic Journal
- Accession number :
- 33577316
- Full Text :
- https://doi.org/10.1021/jacs.0c09213