Journal: Cell reports

Article Title: Functional Genomics Identify Distinct and Overlapping Genes Mediating Resistance to Different Classes of Heterobifunctional Degraders of Oncoproteins

doi: 10.1016/j.celrep.2020.108532

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CRBN and non-targeting control K Stewart’s lab N/A MM.1S CRBN −/− (CRISPR-knockout) W Kaelin’s lab N/A MM.1S-Cas9+ B Ebert’s lab N/A KMS11-Cas9+ Broad Institute, MIT N/A Experimental Models: Organisms/Strains NOD.Cg- Prkdc scid Il2rg tm1Wjl /SzJ The Jackson Laboratory Cat# 005557 Oligonucleotides List of primers – see Table S1 IDT N/A List of sgRNAs for single-gene knock-out – see Table S2 IDT N/A Recombinant DNA lentiCas9-Blast Addgene RRID: Addgene_52962 lentiGuide-Puro Addgene RRID: Addgene_52963 psPAX2 Addgene RRID: Addgene_12260 pMD2.G Addgene RRID: Addgene_12259 GeCKO v2 human sgRNA library Addgene Zhang Lab, Broad Institute of MIT and Harvard Lenti dCAS9-VP64_Blast Addgene RRID: Addgene_61425 pLVX-hyg-sgRNA1 Takara Cat# 632630 Brunello human sgRNA library Addgene RRID: Addgene_73179 Human CRISPR Activation Pooled sgRNA Library ( Calabrese library) Addgene RRID: Addgene_61425 Software and Algorithms MAGeCK Li et al., 2014 https://sourceforge.net/projects/mageck/ PRISM 6 GraphPad https://www.graphpad.com FlowJo V9.7.6 Tree Star https://www.flowjo.com/ Biorender https://www.biorender.com/ Open in a separate window KEY RESOURCES TABLE PROTAC resistance via disruption of rather than adaptation to oncoprotein degradation PROTACs using different E3s/CRLs: resistance via similar pathways but different genes Result of using two PROTACs depends on E3, target, and sequential versus concurrent use E3s essential for and highly expressed in tumor cells are useful for future PROTACs

Techniques: Recombinant, Staining, Cell Viability Assay, Cell Culture, Gel Extraction, Gene Expression, Control, CRISPR, Knock-Out, Activation Assay, Software