Differential substrate degradation by super-degron: EGFP in wild-type mouse cells, PD-1 requires CRBN humanization
Chie Naruse, Ojiro Ishibashi, Masatoshi Ohgushi, Hirohiko Imai, Tomoko Matsuzaki, Xuchi Pan, Tatsuhiko Miyazaki, Yuka Shidahara, Yu Shirakawa, Fumihiro Sugiyama, Masahide Asano
TL;DR
A new method using a super-degron tag and thalidomide analogs can degrade specific proteins in mouse cells, but some proteins require humanized components for effective degradation.
Contribution
The super-degron tag combined with iberdomide enables protein degradation in wild-type mouse cells, with PD-1 degradation requiring CRBN humanization.
Findings
SD-tagged EGFP was degraded in wild-type mouse cells using iberdomide or mezigdomide.
SD-tagged PD-1 required human-type CRBNI391V for degradation in mouse cells.
Iberdomide suppressed tumor growth faster than anti-PD-1 antibodies by degrading PD-1 in vivo.
Abstract
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…
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Taxonomy
TopicsProtein Degradation and Inhibitors · Ubiquitin and proteasome pathways · CAR-T cell therapy research