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C-terminal amides mark proteins for degradation via SCF-FBXO31.

Muhar MF, Farnung J, Cernakova M, Hofmann R, Henneberg LT, Pfleiderer MM, Denoth-Lippuner A, Kalcic F, Nievergelt AS, Peters Al-Bayati M, Sidiropoulos ND, Beier V, Mann M, Jessberger S, Jinek M, Schulman BA, Bode JW, Corn JE
Nature. 2025 Feb;638(8050):519-527. doi: 10.1038/s41586-024-08475-w. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
250247pJC2281 - pHR-SFFV-rtTA3-PGK-BsrExpression of reverse tet transactivator for use in the Tet-On system.
250251pJC2297 - pHR-TRE3G-hSpCas9-NLS-FLAG-2A-Thy1.1Expression of Cas9 with Thy1.1 cell surface marker under a doxycycline responsive promoter. Requires additional expression of transactivator.
250254pJC2722 - pCDNA4TO-CMV-3xFLAG-Halo-AARS1cDNA expression of affinity-tagged AARS1.
250255pJC2723 - pLenti-X1-EF1A-HA-FBXO31-PGK-NeocDNA expression of affinity-tagged FBXO31 (HA-FBXO31).
250256pJC2724 - pLenti-X1-EF1A-HA-FBXO31(∆F-box)-PGK-NeocDNA expression of affinity-tagged FBXO31 (HA-FBXO31(∆F-box)).
250257pJC2725 - pLenti-X1-EF1A-HA-FBXO31(Y309A)-PGK-NeocDNA expression of affinity-tagged FBXO31 (HA-FBXO31(Y309A)).
250258pJC2726 - pLenti-X1-EF1A-HA-FBXO31(I337D)-PGK-NeocDNA expression of affinity-tagged FBXO31 (HA-FBXO31(I337D)).
250259pJC2727 - pLenti-X1-EF1A-HA-FBXO31(T343V)-PGK-NeocDNA expression of affinity-tagged FBXO31 (HA-FBXO31(T343V)).
250260pJC2728 - pLenti-X1-EF1A-HA-FBXO31(D334N)-PGK-NeocDNA expression of affinity-tagged FBXO31 (HA-FBXO31(D334N)).
250263pJC2731 - pHR-EF1A-HA-FBXO31-IRES-GFPcDNA expression of affinity-tagged FBXO31 (HA-FBXO31).
250264pJC2732 - pHR-EF1A-HA-FBXO31(∆F-box)-IRES-GFPcDNA expression of affinity-tagged FBXO31 (HA-FBXO31(∆F-box)).
250265pJC2733 - pHR-EF1A-HA-FBXO31(∆F-box, T343V)-IRES-GFPcDNA expression of affinity-tagged FBXO31 (HA-FBXO31(∆F-box, T343V)).
250266pJC2734 - pHR-EF1A-HA-FBXO31(D334N)-IRES-GFPcDNA expression of affinity-tagged FBXO31 (HA-FBXO31(D334N)).
250267pJC2735 - pHR-EF1A-HA-FBXO31(∆F-box, D334N)-IRES-GFPcDNA expression of affinity-tagged FBXO31 (HA-FBXO31(∆F-box, D334N)).
250268pJC2737 - pHR-EF1A-DD-3xFLAG-FBXO31-IRES-GFPInducible expression of FBXO31 with the shield degron system.
250269pJC2738 - pHR-EF1A-DD-3xFLAG-FBXO31(D334N)-IRES-GFPInducible expression of FBXO31(D334N) with the shield degron system.
250271pJC2740 - pHR-EF1A-HA-FBXO31(D334N)-IRES-mTagBFPcDNA expression of affinity-tagged FBXO31 (HA-FBXO31(D334N))
250272pJC2741 - hPGK-EGFP-CycD1(C-term)-IRES2-mCherry-EF1A-PuroRatiometric protein stability reporter for measuring C-terminal degron activity.
250273pJC2742 - hPGK-EGFP-SUGT1(C-term)-IRES2-mCherry-EF1A-PuroRatiometric protein stability reporter for measuring C-terminal degron activity.
250274pJC2743 - hPGK-EGFP-MTFR1L(C-term)-IRES2-mCherry-EF1A-PuroRatiometric protein stability reporter for measuring C-terminal degron activity.
250275pJC2744 - hPGK-EGFP-GOPC(C-term)-IRES2-mCherry-EF1A-PuroRatiometric protein stability reporter for measuring C-terminal degron activity.
250277pJC2746 - pLC-U6-spacer-modified-tracer-EFS-eBFP2-P2A-PuroEmpty backbone for cloning and stable sgRNA expression.

Antibodies from Article