KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal FUS/TLS antibody | Santa Cruz Biotech | sc-47711 |
| eIF3b antibody | Santa Cruz Biotech | sc-16377 |
| Mouse monoclonal G3BP1 antibody | Santa Cruz Biotech | sc-365338 |
| Rabbit monoclonal MAR/PAR antibody | Cell Signaling Tech | 83732 |
| Rabbit polyclonal TNKS antibody | Santa Cruz Biotech | sc-8337 |
| Monoclonal Halo-Tag antibody | Promega | G9211 |
| Pan-ADP-ribose binding reagent | Millipore | MABE1016 |
| Mouse anti-alpha Tubulin antibody [DM1α] | Abcam | ab7291 |
| Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 | Invitrogen | A21202 |
| Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) | Abcam | ab150077 |
| Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 | Invitrogen | A27012 |
| Goat anti-Mouse IgG (H+L) Irdye 800 CW Secondary Antibody | Li-Cor | 926-32210 |
| Goat anti-Rabbit IgG (H+L) Irdye 680 RD Secondary Antibody | Li-Cor | 926-68071 |
| Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP | Thermo Scientific | 31430 |
| Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP | Thermo Scientific | 31460 |
| Revert 700 Total Protein Stain | Li-Cor | 926-11011 |
| Bacterial and Virus Strains | ||
| E. Coli BL21(DE3) Chemically Competent Cells | Millipore Sigma | CMC0014-20X40UL |
| BL21-CodonPlus (DE3)-RIL Competent Cells | Agilent | 230245 |
| NEB® Turbo Competent E. coli (High Efficiency) | New England Biolabs | C2984H |
| Chemicals, Peptides, and Recombinant Proteins | ||
| AcTEV Protease | Fisher Scientific | 12-575-015 |
| Ribonuclease A from bovine pancreas | Millipore Sigma | R6513 |
| cOmplete™ Protease Inhibitor Cocktail | Roche | 11697498001 |
| Glucose Oxidase from Aspergillus niger | Sigma-Aldrich | G2133 |
| Catalase from bovine liver | Sigma-Aldrich | C3155 |
| IGEPAL® CA-630 | Sigma-Aldrich | I8896 |
| Alexa Fluor™ 488 TFP ester | Thermo Fisher | A37563 |
| Cy3 NHS Ester | Cytiva | PA13101 |
| Cy5 NHS Ester | Cytiva | PA15100 |
| Janelia Fluor® 549 HaloTag® Ligand | Promega | GA1110 |
| Rnase Inhibitor, Murine | New England Biolabs | M0314L |
| 1,6-Hexanediol,99% | Sigma-Aldrich | 240117-50G |
| Lipofectamine™ 3000 Transfection Reagent | Thermo Scientific | L3000001 |
| Sodium (meta)arsenite (>90%) | Millipore Sigma | S7400-100G |
| Fibronectin bovine plasma | Millipore Sigma | F1141-1MG |
| Poly-D-lysine | R&D Systems | 3439-200-01 |
| Geneticin™ Selective Antibiotic (G418 Sulfate) | Thermo Scientific | 10131035 |
| NP40 Cell Lysis Buffer | Thermo Scientific | FNN0021 |
| Pierce™ ECL Western Blotting Substrate | Thermo Scientific | 32109 |
| Olaparib | Selleckchem | S1060 |
| G007-LK | Sigma-Aldrich | 504907 |
| IWR-1 | Cayman Chemical | 13659 |
| PDD 00017273 | MedChemExpress | HY-108360 |
| Alkyne linker: (2-[2-(2-Propynyloxy)ethoxy]ethylamine) CAS# 944561-44-8 | TCI | P2225 |
| dATP-Cy3= N6-(6-Aminohexyl)-dATP-Cy3 | Jena Bioscience | NU-835-CY3 |
| EDAC, Hydrochloride | Sigma-Aldrich | 25952-53-8 |
| Tris(3-hydroxypropyltriazolylmethyl)amine | Sigma-Aldrich | 760952-88-3 |
| Experimental Models: Cell Lines | ||
| Human: SH-SY5Y | ATCC | CRL-2266 |
| Human: WT-FUS-Halo SH-SY5Y (GenR) | This paper | N/A |
| Oligonucleotides | ||
| Biotin-18mer: 5’- /biotin/rUrGrG rCrGrA rCrGrG rCrArG rCrGrA rGrGrC/3AmMO/ −3’ | Niaki et al., 2020; IDT | N/A |
| U50-18mer: 5’- /5AmMC6/rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrG rCrCrU rCrGrC rUrGrC rCrGrU rCrGrC rCrA −3’ | Niaki et al., 2020; IDT | N/A |
| U40: 5’- rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rU/3AmMO/ −3’ | Niaki et al., 2020; IDT | N/A |
| Bio-U40: 5’- rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rUrUrU rU/Bio/ −3’ | IDT | N/A |
| DNA-18-mer: 5’- TGG CGA CGG CAG CGA GGC −3’ | IDT | N/A |
| Bio-DNA-18-mer: 5’- /Cy5/GCC TCG CTG CCG TCG CCA/biotin/ −3’ | IDT | N/A |
| PARP1-shRNA-1-F: 5’-rUrGrArCrUrUrGrGrArArGrUrCrArUrCrGrArUrArUrCrUTT-3’ | IDT | hs.Ri.PARP1.13.1 |
| PARP1-shRNA-1-R: 5’-rArArArGrArUrArUrCrGrArUrGrArCrUrUrCrCrArArGrUrCrArUrA-3’ | IDT | hs.Ri.PARP1.13.1 |
| PARP1-shRNA-2-F: 5’-rCrUrGrArArGrGrArGrCrUrArCrUrCrArUrCrUrUrCrArACA-3’ | IDT | hs.Ri.PARP1.13.2 |
| PARP1-shRNA-2-R: 5’-rUrGrUrUrGrArArGrArUrGrArGrUrArGrCrUrCrCrUrUrCrArGrGrU-3’ | IDT | hs.Ri.PARP1.13.2 |
| PARP1-shRNA-3-F: 5’-rGrGrArArGrGrUrArUrCrArArCrArArArUrCrUrGrArArAAG | IDT | hs.Ri.PARP1.13.6 |
| PARP1-shRNA-3-R: 5’-rCrUrUrUrUrCrArGrArUrUrUrGrUrUrGrArUrArCrCrUrUrCrCrUrC-3’ | IDT | hs.Ri.PARP1.13.6 |
| PARP5a-shRNA-1-F: 5’-rGrUrCrArCrArGrArArCrUrGrCrUrArCrUrArArArGrCrATG-3’ | IDT | hs.Ri.TNKS.13.1 |
| PARP5a-shRNA-1-R: 5’-rrArUrGrCrUrUrUrArGrUrArGrCrArGrUrUrCrUrGrUrGrArCrUrU-3’ | IDT | hs.Ri.TNKS.13.1 |
| PARP5a-shRNA-2-F: 5’- rArUrGrArArUrArUrCrArGrCrCrArArUrUrUrCrUrArArAAA-3’ | IDT | hs.Ri.TNKS.13.2 |
| PARP5a-shRNA-2-R: 5’-rUrUrUrUrUrArGrArArArUrUrGrGrCrUrGrArUrArUrUrCrArUrGrU-3’ | IDT | hs.Ri.TNKS.13.2 |
| PARP5a-shRNA-3-F: 5’-rGrArCrArGrArArUrUrGUrUrArCrUrUrArGrArArArArGGA-3’ | IDT | hs.Ri.TNKS.13.5 |
| PARP5a-shRNA-3-R: 5’-rUrCrCrUrUrUrUrCrUrArArGrUrArArCrArArUrUrCrUrGrUrCrArC-3’ | IDT | hs.Ri.TNKS.13.5 |
| Recombinant DNA | ||
| pTHMT/FUSWT (encoding 6xHis-MBP-FUS-WT) | Niaki et al., 2020 | N/A |
| pTHMT/FUSR244C (encoding 6xHis-MBP-FUS-R244C) | Niaki et al., 2020 | N/A |
| pTHMT/FUSG156E (encoding 6xHis-MBP-FUS-G156E) | Niaki et al., 2020 | N/A |
| pTHMT/FUSY>S (encoding 6xHis-MBP-FUS(Y>S)) | Genscript | N/A |
| pTHMT/FUSR>G (encoding 6xHis-MBP-FUS(R>G)) | Genscript | N/A |
| Super piggyBac Transposase expression vector | Systems Biosciences | PB210PA-1 |
| pFUSWT-Halo_piggybac-EF1-Halo | Rhine et al., 2020a | N/A |
| pET24a-PARP5a (1093-1327) | Tan et al., 2012 | N/A |
| pGFP-cPARG102 | Leung et al., 2011 | N/A |
| pSAT1-PARG (318-976) | This paper | N/A |
| GST-TEV-Kapβ2 | Guo et al., 2018 | N/A |
| Software and Algorithms | ||
| Matlab | Mathworks | N/A |
| NIS-Elements Ar Package | Nikon Inc. | N/A |
| Adobe Photoshop CC | Adobe (https://www.adobe.com/products/photoshop.html ) | N/A |
| Prism 8 | GraphPad (https://www.graphpad.com/scientific-software/prism/ ) | N/A |
| ImageJ (Fiji) | NIH (https://imagej.nih.gov/ij/ ) | N/A |
| ImageStudio 5.2 | Li-Cor Biosciences | N/A |
| Zen Blue | Zeiss | N/A |
| Matlab code | This paper | DOI: 10.5281/zenodo.5866686 |
| Other | ||
| Nunc™ Lab-Tek™ Chambered Coverglass | Thermo Scientific | 155361 |
| Nunc™ Lab-Tek™ II Chamber Slide™ System | Thermo Scientific | 154526PK |
| Superdex™ 200 Increase 10/300 GL | Cytiva | 28990944 |
| HisTrap™ Fast Flow Crude | Cytiva | 17528601 |
| HisPur™ Ni-NTA Resin | Thermo Scientific | 88221 |
| Glutathione Sepharose 4 Fast Flow, 100 mL | Cytiva | 17513202 |
| HiTrap™ Q HP | Cytiva | 17115401 |
| Streptavidin Coated Polystyrene Particles | Spherotech | SVP-40-5 |
| DNA Retardation Gels (6%) | Invitrogen | EC63655BOX |
| Any kD™ Mini-PROTEAN® TGX™ Precast Protein Gels, 15-well, 15 μL | Bio Rad | 4569036 |
| Nitrocellulose membranes, roll, pore size 0.45 μm | VWR | 10-6000-02 |
| Amicon™ Ultra-0.5 Centrifugal Filter Unit (10K MWCO) | Millipore Sigma | UFC501096 |
| Amicon™ Ultra-15 Centrifugal Filter Unit (10K MWCO) | Millipore Sigma | UFC901024 |
| Zeba™ Spin Desalting Columns, 7K MWCO, 0.5 mL | Thermo Scientific | 89883 |
| Corning™ 96-Well, Non-Treated, Flat-Bottom, Half-Area Microplate | Fisher Scientific | 07-200-840 |
| High Pure miRNA Isolation Kit | Roche | 5080576001 |
| HiTrap Desalting | Cytiva | 17140801 |
| Monarch PCR & DNA Cleanup Kit | New England Biolabs | T1030S |