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Protein degradation by TRIM-Away, a novel tool learned from innate immunity
- 2. Methods for targeted disruption of protein function DNA level: Knockout by TALEN/CRISPER-Cas9, etc. mRNA level: RNAi knockdown Protein level: drugs OR induced protein degradation Backgrounds Buckley and Crews, 2014
- 3. Key points in exploiting UPS to induce specific degradation of protein Backgrounds UPS Engagement Target binding Small molecule Antibody Known PPI E3 fusion E3 recruiting Degron
- 4. PROTAC molecule recruit E3 ligase to its target to induce target degradation Backgrounds Burslem and Crews, 2017
- 5. Basic structure of conventional antibody Backgrounds Google Search
- 6. From Conventional antibody to nanobody Backgrounds Google Search
- 7. Fusion of antibody to E3 ligase could efficiently degrade target proteins Backgrounds Caussinus et al., 2011 Portnoff et al., 2014
- 8. TRIM21 is an cytosolic IgG receptor in innate immunity Google Backgrounds
- 9. TRIM21 is an cytosolic IgG receptor in innate immunity MRC lab, UK Backgrounds
- 10. Schematic of TRIM-AWAY Approach Results
- 11. TRIM-Away worked efficiently in degrading free GFP Results
- 12. GFP-degradation by TRIM-Away was mediated by proteasome Results
- 13. TRIM-Away could degrade proteins localized to different cellular regions Results
- 14. TRIM-Away could degrade proteins localized to different cellular regions Results
- 15. Nucleus-shuttling nanobody help TRIM- Away degrade nucleus-retained proteins Results
- 16. TRIM-Away specifically degrade endogenous proteins in primary oocyte Results
- 17. TRIM-Away specifically degrade endogenous proteins in primary oocyte Results
- 18. TRIM-Away could specifically degrade certain isoform of target protein Results
- 19. TRIM-Away specifically degrade long- life proteins Results
- 20. TRIM-Away specifically degrade long- life proteins Results
- 21. TRIM-Away specifically degrade long- life proteins Results
- 22. Antibody could be introduced into cell population by electroporation Results
- 23. TRIM-Away of pericentrin by antibody electroporation Results
- 24. Selective Trim-Away of Signaling Pathway Components Results
- 25. Selective Trim-Away of Signaling Pathway Components Results
- 26. TRIM-Away could work in various unmodified cell lines Results
- 27. TRIM-Away could work in primary human macrophages Results
- 28. TRIM-Away could work in primary human macrophages Results
- 29. TRIM21 concomitantly degrades with target proteins Results
- 30. TRIM21 and antibody need to excess target to guarantee TRIM-Away Results
- 31. Duration of target depletion by TRIM- Away Results
- 32. Remaining challenges The expression level of TRIM21 is a limiting factor Microinjection/electroporation of TRIM-Away is too complicated for broad therapeutic applications Discussion
- 33. Take-home Messages--Highlights Trim-Away is a widely applicable method to degrade endogenous proteins Target proteins do not need to be modified before degradation Proteins are degraded within minutes of application Trim-Away allows efficient protein depletion in primary human cells Conclusion
Editor's Notes
- I think all the strategies developed till now provide their answers to one or both of these two issues, i.e., how to bind their targets and how to engage the UPS.
- Trim21 recognises antibodies which enter the cell attached to viruses shown in polyhedron. It binds to these antibodies, tags the antibody-virus-complex as ‘garbage’, and hands it over to the cell’s ‘garbage chute’, the proteasome.
- Trim21 recognises antibodies which enter the cell attached to viruses shown in polyhedron. It binds to these antibodies, tags the antibody-virus-complex as ‘garbage’, and hands it over to the cell’s ‘garbage chute’, the proteasome. 你有张良计，我有过墙梯
- Abcam ab6556 rabbit polyclonal GFP antibody
- Phenotype！！！