KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-p53 | Santa Cruz Biotechnology | Cat# sc-126; RRID: AB_628082 |
| mouse monoclonal anti-MDM2 (Ab-1) | Millipore | Cat# OP46T-10UG; RRID: AB_564805 |
| rabbit anti-pChk2 (T68) | Cell Signaling Technology | Cat# 2661; RRID: AB_331479 |
| rabbit anti-pChk1 (S317) | Cell Signaling Technology | Cat# 2344; RRID: AB_331448 |
| mouse anti-tubulin (E7) | Developmental Studies Hybridoma Bank | N/A |
| mouse monoclonal anti-β-Actin (AC-74) | Sigma-Aldrich | Cat# A2228; RRID: AB_476697 |
| HRP donkey anti-rabbit | VWR | Cat# 95017–556 |
| HRP goat anti-mouse | Thermo-Fisher | Cat# G21040; RRID: AB_2536527 |
| IRDye® 680RD donkey anti-rabbit | Li-COR | Cat# 925–68073; RRID: AB_2716687 |
| IRDye® 800CW goat anti-rabbit | Li-COR | Cat# 926–32211; RRID: AB_621843 |
| IRDye® 680RD goat anti-mouse | Li-COR | Cat# 926–68070; RRID: AB_10956588 |
| IRDye® 800CW donkey anti-mouse | Li-COR | Cat# 925–32212; RRID: AB_2716622 |
| Bacterial and Virus Strains | ||
| lentiCRISPR v2 | Feng Zhang | Addgene plasmid # 52961; http://addge.neorg/52961; RRID:Addgene_52961 |
| leniGuide-puro | Feng Zhang | Addgene plasmid # 52963; http://addgene.org/52963; RRID:Addgene_52963 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| ATM inhibitor (KU55933) | Sigma-Aldrich | Cat# SML1109 |
| ATR inhibitor (VE822) | Cayman Chemical | Cat# 24198 |
| Caspase 2 inhibitor (Z-VDVAD-FMK) | R&D Systems | Cat# 2165 (discontinued) |
| Caspase 2 inhibitor (NH-23-C2) | Poreba et al., 2019 | N/A |
| Doxycycline | Sigma-Aldrich | Cat# D9891 |
| Ribociclib | MedChem Express | Cat# HY-15777 |
| SMARTpool siGenome CDKN1A (p21) | Dharmacon | Cat M-003471–00-0005 Lot 140909 |
| Critical Commercial Assays | ||
| RNAi MAX transfection reagent | Thermo Fisher Scientific | Cat# 13778075 |
| Experimental Models: Cell Lines | ||
| Human: MCF7 pMT-p53-mVenus | Purvis et al., 2012 | N/A |
| Human: MCF7 pMT-p53-mVenus pCW22TRE-PIDD-mCer | This work | N/A |
| Human: MCF7 pMT-p53-mVenus PIDD1 KO | This work | N/A |
| Human: MCF7 pUb-p53-mVenus | Stewart-Ornstein and Lahav, 2017 | N/A |
| Human: RPE p21-mKate2 + pUBp-p53-mNeonGreen | Reyes et al., 2018 | N/A |
| Human: A549 pUb-p53-mNeonGreen H2B-mCherry | Stewart-Orenstein et al., 2017 | N/A |
| Oligonucleotides | ||
| PIDD1 siRNA sequences - sense | rCrArGrArCrUrGrUrUrCrCrUrG rArCrCrUrCrArGrArUrU |
|
| PIDD1 siRNA sequences - antisense | rCrArGrArCrUrGrUrUrCrCrUr GrArCrCrUrCrArGrArUrU |
|
| PIDD1 sgRNA forward | CACCGGCGGGAAACC ACAAGGAACC |
|
| PIDD1 sgRNA reverse | AAACGGTTCCTTGTG GTTTCCCGC |
|
| Recombinant DNA | ||
| pCW22TRE-PIDD1-mCer | This study | N/A |
| PIDD1-sgRNA-puro | This study | N/A |
| Software and Algorithms | ||
| P53Cinema Single Cell Tracking Software | Reyes et al., 2018 | https://github.com/balvahal/p53CinemaManual |
| Model for p53 regulation through MDM2 degradation by the PIDDosome | This study | https://github.com/MTsabar/PIDDosome/tree/PIDDosome |