Uncovering receptor-ligand interactions using a high-avidity CRISPR activation screening platform
The majority of clinically approved drugs target proteins that are secreted or cell surface bound. However, further advances in this area have been hindered by the challenging nature of receptor deorphanization, as there are still many secreted and cell-bound proteins with unknown binding partners. Here, we developed an advanced screening platform that combines CRISPR-CAS9 guide-mediated gene activation (CRISPRa) and high-avidity bead-based selection. The CRISPRa platform incorporates serial enrichment and flow cytometry-based monitoring, resulting in substantially improved screening sensitivity for well-known yet weak interactions of the checkpoint inhibitor family. Our approach has successfully revealed that siglec-4 exerts regulatory control over T cell activation through a low affinity trans-interaction with the costimulatory receptor 4-1BB. Our highly efficient screening platform holds great promise for identifying extracellular interactions of uncharacterized receptor-ligand partners, which is essential to develop next-generation therapeutics, including additional immune checkpoint inhibitors.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:10 |
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| Enthalten in: |
Science advances - 10(2024), 7 vom: 16. Feb., Seite eadj2445 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Yang, Liping [VerfasserIn] |
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| Links: |
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| Themen: |
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| Anmerkungen: |
Date Completed 16.02.2024 Date Revised 01.03.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1126/sciadv.adj2445 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM368443418 |
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| 500 | |a Citation Status MEDLINE | ||
| 520 | |a The majority of clinically approved drugs target proteins that are secreted or cell surface bound. However, further advances in this area have been hindered by the challenging nature of receptor deorphanization, as there are still many secreted and cell-bound proteins with unknown binding partners. Here, we developed an advanced screening platform that combines CRISPR-CAS9 guide-mediated gene activation (CRISPRa) and high-avidity bead-based selection. The CRISPRa platform incorporates serial enrichment and flow cytometry-based monitoring, resulting in substantially improved screening sensitivity for well-known yet weak interactions of the checkpoint inhibitor family. Our approach has successfully revealed that siglec-4 exerts regulatory control over T cell activation through a low affinity trans-interaction with the costimulatory receptor 4-1BB. Our highly efficient screening platform holds great promise for identifying extracellular interactions of uncharacterized receptor-ligand partners, which is essential to develop next-generation therapeutics, including additional immune checkpoint inhibitors | ||
| 650 | 4 | |a Journal Article | |
| 650 | 7 | |a Ligands |2 NLM | |
| 650 | 7 | |a Membrane Proteins |2 NLM | |
| 700 | 1 | |a Sheets, Timothy P |e verfasserin |4 aut | |
| 700 | 1 | |a Feng, Yang |e verfasserin |4 aut | |
| 700 | 1 | |a Yu, Guojun |e verfasserin |4 aut | |
| 700 | 1 | |a Bajgain, Pradip |e verfasserin |4 aut | |
| 700 | 1 | |a Hsu, Kuo-Sheng |e verfasserin |4 aut | |
| 700 | 1 | |a So, Daeho |e verfasserin |4 aut | |
| 700 | 1 | |a Seaman, Steven |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Jaewon |e verfasserin |4 aut | |
| 700 | 1 | |a Lin, Ling |e verfasserin |4 aut | |
| 700 | 1 | |a Evans, Christine N |e verfasserin |4 aut | |
| 700 | 1 | |a Guest, Mary R |e verfasserin |4 aut | |
| 700 | 1 | |a Chari, Raj |e verfasserin |4 aut | |
| 700 | 1 | |a St Croix, Brad |e verfasserin |4 aut | |
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