Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanization
Summary: Protein knockdown using a zinc finger degron tag and thalidomide analogs was previously considered ineffective in mouse cells. However, using EGFP as an indicator, we found that a super-degron tag (SD) enables degradation in mouse cells when combined with iberdomide or mezigdomide. While SD...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-07-01
|
| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004225012532 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850116654004961280 |
|---|---|
| author | Chie Naruse Ojiro Ishibashi Masatoshi Ohgushi Hirohiko Imai Tomoko Matsuzaki Xuchi Pan Tatsuhiko Miyazaki Yuka Shidahara Yu Shirakawa Fumihiro Sugiyama Masahide Asano |
| author_facet | Chie Naruse Ojiro Ishibashi Masatoshi Ohgushi Hirohiko Imai Tomoko Matsuzaki Xuchi Pan Tatsuhiko Miyazaki Yuka Shidahara Yu Shirakawa Fumihiro Sugiyama Masahide Asano |
| author_sort | Chie Naruse |
| collection | DOAJ |
| description | Summary: Protein knockdown using a zinc finger degron tag and thalidomide analogs was previously considered ineffective in mouse cells. However, using EGFP as an indicator, we found that a super-degron tag (SD) enables degradation in mouse cells when combined with iberdomide or mezigdomide. While SD-tagged EGFP was degraded in wild-type mouse cells, SD-tagged PD-1 required human-type CEREBLON (CRBNI391V) for degradation. In mice with CRBNI391V, endogenous PD-1 tagged with SD was efficiently degraded in T cells both in vitro and in vivo. Compared with anti-PD-1 antibody treatment, the degradation of PD-1 led to more rapid activation of CD8+ T cells. Moreover, pomalidomide that crosses the brain-blood barrier reduced PD-1 expression in the brain. These results suggest that SD and thalidomide analogs can be used for in vitro and in vivo protein knockdown in mice, although some conditional settings are required. |
| format | Article |
| id | doaj-art-bcc99651c2e449b897a764ca6ce9a3f1 |
| institution | OA Journals |
| issn | 2589-0042 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj-art-bcc99651c2e449b897a764ca6ce9a3f12025-08-20T02:36:16ZengElsevieriScience2589-00422025-07-0128711299210.1016/j.isci.2025.112992Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanizationChie Naruse0Ojiro Ishibashi1Masatoshi Ohgushi2Hirohiko Imai3Tomoko Matsuzaki4Xuchi Pan5Tatsuhiko Miyazaki6Yuka Shidahara7Yu Shirakawa8Fumihiro Sugiyama9Masahide Asano10Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan; Corresponding authorInstitute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, JapanLaboratory of Developmental Systems, Department of Biosystems Science, Institute for Life and Medical Sciences, Kyoto University, Kyoto 606-8507, JapanDepartment of Informatics, Graduate School of informatics, Kyoto University, Kyoto 606-8501, Japan; Innovation Research Center for Quantum Medicine, Gifu University Hospital, Gifu 501-1194, JapanInstitute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, JapanInstitute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, JapanDepartment of Pathology, Gifu University Hospital, Gifu 501-1194, JapanBioscience Business Division, KAC Co., Ltd. Ritto, Shiga 520-3001, JapanBioscience Business Division, KAC Co., Ltd. Ritto, Shiga 520-3001, JapanLaborarory Animal Resource Center, Transborder Medical Research Center, Institute of Medicine, University of Tsukuba, Tsukuba 305-8575, JapanInstitute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan; Corresponding authorSummary: Protein knockdown using a zinc finger degron tag and thalidomide analogs was previously considered ineffective in mouse cells. However, using EGFP as an indicator, we found that a super-degron tag (SD) enables degradation in mouse cells when combined with iberdomide or mezigdomide. While SD-tagged EGFP was degraded in wild-type mouse cells, SD-tagged PD-1 required human-type CEREBLON (CRBNI391V) for degradation. In mice with CRBNI391V, endogenous PD-1 tagged with SD was efficiently degraded in T cells both in vitro and in vivo. Compared with anti-PD-1 antibody treatment, the degradation of PD-1 led to more rapid activation of CD8+ T cells. Moreover, pomalidomide that crosses the brain-blood barrier reduced PD-1 expression in the brain. These results suggest that SD and thalidomide analogs can be used for in vitro and in vivo protein knockdown in mice, although some conditional settings are required.http://www.sciencedirect.com/science/article/pii/S2589004225012532Natural sciencesBiological sciencesImmunology |
| spellingShingle | Chie Naruse Ojiro Ishibashi Masatoshi Ohgushi Hirohiko Imai Tomoko Matsuzaki Xuchi Pan Tatsuhiko Miyazaki Yuka Shidahara Yu Shirakawa Fumihiro Sugiyama Masahide Asano Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanization iScience Natural sciences Biological sciences Immunology |
| title | Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanization |
| title_full | Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanization |
| title_fullStr | Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanization |
| title_full_unstemmed | Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanization |
| title_short | Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanization |
| title_sort | differential substrate degradation by super degron egfp in wild type mouse cells pd 1 requires crbn humanization |
| topic | Natural sciences Biological sciences Immunology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004225012532 |
| work_keys_str_mv | AT chienaruse differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT ojiroishibashi differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT masatoshiohgushi differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT hirohikoimai differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT tomokomatsuzaki differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT xuchipan differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT tatsuhikomiyazaki differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT yukashidahara differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT yushirakawa differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT fumihirosugiyama differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization AT masahideasano differentialsubstratedegradationbysuperdegronegfpinwildtypemousecellspd1requirescrbnhumanization |