guest ::
| Home > Publications database > Luciferase- and HaloTag-based reporter assays to measure small-molecule-induced degradation pathway in living cells. > print |
| Home > Publications database > Luciferase- and HaloTag-based reporter assays to measure small-molecule-induced degradation pathway in living cells. > print |
| 001 | 289592 | ||
| 005 | 20250401141726.0 | ||
| 024 | 7 | _ | |a 10.1038/s41596-024-00979-z |2 doi |
| 024 | 7 | _ | |a pmid:38637703 |2 pmid |
| 024 | 7 | _ | |a 1754-2189 |2 ISSN |
| 024 | 7 | _ | |a 1750-2799 |2 ISSN |
| 024 | 7 | _ | |a altmetric:162632051 |2 altmetric |
| 037 | _ | _ | |a DKFZ-2024-00825 |
| 041 | _ | _ | |a English |
| 082 | _ | _ | |a 610 |
| 100 | 1 | _ | |a Schwalm, Martin P |0 0000-0002-1252-1829 |b 0 |
| 245 | _ | _ | |a Luciferase- and HaloTag-based reporter assays to measure small-molecule-induced degradation pathway in living cells. |
| 260 | _ | _ | |a Basingstoke |c 2024 |b Nature Publishing Group |
| 336 | 7 | _ | |a article |2 DRIVER |
| 336 | 7 | _ | |a Output Types/Journal article |2 DataCite |
| 336 | 7 | _ | |a Journal Article |b journal |m journal |0 PUB:(DE-HGF)16 |s 1723034180_32361 |2 PUB:(DE-HGF) |x Review Article |
| 336 | 7 | _ | |a ARTICLE |2 BibTeX |
| 336 | 7 | _ | |a JOURNAL_ARTICLE |2 ORCID |
| 336 | 7 | _ | |a Journal Article |0 0 |2 EndNote |
| 500 | _ | _ | |a 2024 Aug;19(8):2317-2357 |
| 520 | _ | _ | |a The rational development of small-molecule degraders (e.g., proteolysis targeting chimeras) remains a challenge as the rate-limiting steps that determine degrader efficiency are largely unknown. Standard methods in the field of targeted protein degradation mostly rely on classical, low-throughput endpoint assays such as western blots or quantitative proteomics. Here we applied NanoLuciferase- and HaloTag-based screening technologies to determine the kinetics and stability of small-molecule-induced ternary complex formation between a protein of interest and a selected E3 ligase. A collection of live-cell assays were designed to probe the most critical steps of the degradation process while minimizing the number of required expression constructs, making the proposed assay pipeline flexible and adaptable to the requirements of the users. This approach evaluates the underlying mechanism of selective target degraders and reveals the exact characteristics of the developed degrader molecules in living cells. The protocol allows scientists trained in basic cell culture and molecular biology to carry out small-molecule proximity-inducer screening via tracking of the ternary complex formation within 2 weeks of establishment, while degrader screening using the HiBiT system requires a CRISPR-Cas9 engineered cell line whose generation can take up to 3 months. After cell-line generation, degrader screening and validation can be carried out in high-throughput manner within days. |
| 536 | _ | _ | |a 899 - ohne Topic (POF4-899) |0 G:(DE-HGF)POF4-899 |c POF4-899 |f POF IV |x 0 |
| 588 | _ | _ | |a Dataset connected to CrossRef, PubMed, , Journals: inrepo02.dkfz.de |
| 700 | 1 | _ | |a Saxena, Krishna |0 0000-0002-2237-5378 |b 1 |
| 700 | 1 | _ | |a Müller, Susanne |0 0000-0003-2402-4157 |b 2 |
| 700 | 1 | _ | |a Knapp, Stefan |0 P:(DE-HGF)0 |b 3 |
| 773 | _ | _ | |a 10.1038/s41596-024-00979-z |0 PERI:(DE-600)2244966-8 |n 8 |p 2317-2357 |t Nature protocols |v 19 |y 2024 |x 1754-2189 |
| 856 | 4 | _ | |u https://inrepo02.dkfz.de/record/289592/files/s41596-024-00979-z.pdf |
| 856 | 4 | _ | |u https://inrepo02.dkfz.de/record/289592/files/s41596-024-00979-z.pdf?subformat=pdfa |x pdfa |
| 909 | C | O | |p VDB |o oai:inrepo02.dkfz.de:289592 |
| 910 | 1 | _ | |a Deutsches Krebsforschungszentrum |0 I:(DE-588b)2036810-0 |k DKFZ |b 0 |6 0000-0002-1252-1829 |
| 910 | 1 | _ | |a Deutsches Krebsforschungszentrum |0 I:(DE-588b)2036810-0 |k DKFZ |b 3 |6 P:(DE-HGF)0 |
| 913 | 1 | _ | |a DE-HGF |b Programmungebundene Forschung |l ohne Programm |1 G:(DE-HGF)POF4-890 |0 G:(DE-HGF)POF4-899 |3 G:(DE-HGF)POF4 |2 G:(DE-HGF)POF4-800 |4 G:(DE-HGF)POF |v ohne Topic |x 0 |
| 914 | 1 | _ | |y 2024 |
| 915 | _ | _ | |a Nationallizenz |0 StatID:(DE-HGF)0420 |2 StatID |d 2023-10-25 |w ger |
| 915 | _ | _ | |a DEAL Nature |0 StatID:(DE-HGF)3003 |2 StatID |d 2023-10-25 |w ger |
| 915 | _ | _ | |a DBCoverage |0 StatID:(DE-HGF)0300 |2 StatID |b Medline |d 2023-10-25 |
| 915 | _ | _ | |a DBCoverage |0 StatID:(DE-HGF)0199 |2 StatID |b Clarivate Analytics Master Journal List |d 2023-10-25 |
| 915 | _ | _ | |a WoS |0 StatID:(DE-HGF)0113 |2 StatID |b Science Citation Index Expanded |d 2023-10-25 |
| 915 | _ | _ | |a DBCoverage |0 StatID:(DE-HGF)0150 |2 StatID |b Web of Science Core Collection |d 2023-10-25 |
| 915 | _ | _ | |a DBCoverage |0 StatID:(DE-HGF)1030 |2 StatID |b Current Contents - Life Sciences |d 2023-10-25 |
| 915 | _ | _ | |a DBCoverage |0 StatID:(DE-HGF)0160 |2 StatID |b Essential Science Indicators |d 2023-10-25 |
| 915 | _ | _ | |a DBCoverage |0 StatID:(DE-HGF)0200 |2 StatID |b SCOPUS |d 2023-10-25 |
| 920 | 1 | _ | |0 I:(DE-He78)FM01-20160331 |k FM01 |l DKTK Koordinierungsstelle Frankfurt |x 0 |
| 980 | _ | _ | |a journal |
| 980 | _ | _ | |a VDB |
| 980 | _ | _ | |a I:(DE-He78)FM01-20160331 |
| 980 | _ | _ | |a UNRESTRICTED |
| Library | Collection | CLSMajor | CLSMinor | Language | Author |
|---|