Reagents and tools table
| Reagent/resource | Reference or source | Identifier or catalogue number |
|---|---|---|
| Experimental models | ||
| NIKS | Allen-Hoffman et al J Invest Dermatol 114(3):444—55 (2000) | Cellosaurus Accession CVCL_A1LW |
| 3T3-J2 | Rheinwald J.G., Green H. Cell 6:331–343 (1975) | Cellosaurus Accession CVCL_W667 |
| Normal primary human keratinocytes from pooled adult donors (NHEK) | Sigma-Aldrich | C-12006 |
| Recombinant DNA | ||
| pCR Blunt II-TOPO | Thermo Fisher | 450245 |
| pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A) | Addgene (Cong et al Science 15;339(6121):819-23 (2013)) | 42335 |
| pEGFP-N1-flpo | This study | |
| APOBEC3A targeting plasmid (pBSA3A-del) | This study | |
| pRH3097-A3A (for qRT-PCR standard) | Reuben Harris / Refsland et al Nucl Acids Res 38(13):4274-84 (2010) | |
| pCMV4-APOBEC3B (for qRT-PCR standard) | Mike Malim (Kings College London, UK) | |
| Antibodies | ||
| APOBEC3A (rabbit monoclonal UMN-13) | In-house (Reuben Harris) / Naumann et al Int. J. Mol. Sci. 24(11), 9305 (2023) | |
| APOBEC3A/B/G (rabbit monoclonal 5210-87-13) | Cell Signaling Technology / Brown et al Antibodies 10;8(3):47 (2019)) | 81001 |
| Oligonucleotides and other sequence-based reagents | ||
| Primers and probes used for digital PCR, qRT-PCR and/or plasmid construction to generate standard curves. |
APOBEC3A, APOBEC3B and TBP qRT-PCR primers: Refsland et al Nucl Acids Res 38(13):4274-84 (2010) DDOST primers and probes: Oh and Buisson STAR Protocols 3(1)101148 (2022) All other primers, this study All primers and probes were purchased from Integrated DNA Technologies. |
Table 1 |
| SilencerSelect Non-targeting siRNA (NC#1) | Thermo Fisher | 4390843 |
| SilencerSelect GRHL3 siRNA #1 | Thermo Fisher | s33753 |
| SilencerSelect GRHL3 siRNA #2 | Thermo Fisher | s33754 |
| Chemicals, Enzymes and other reagents | ||
| Inhibitors: | ||
| Afatinib | Fisher Scientific | 16463748 |
| Everolimus | Fluka Analyticals | 57-55-6 |
| GÖ6983 | Sigma-Aldrich | G1918 |
| MK2206 | Selleck Chemicals | S1078 |
| Phorbol 12-myristate 13-acetate (PMA) | Stem Cell Technologies | 74042 |
| Pictilosib (GDC0941) | Selleck Chemicals | S1065 |
| Ravoxertinib (GDC0994) | Selleck Chemicals | S7554 |
| Trametinib (GSK1120212) | Selleck Chemicals | NC0991754 |
| Cell culture: | ||
| Adenine | Sigma-Aldrich | A2786 |
| Cholera Toxin | Merck-Millipore | 227036 |
| Dulbecco’s modified eagle medium (DMEM) with 4.5 g/L glucose, 3.7 g/L NaHCO3, with l-glutamine, without sodium pyruvate | PAN Biotech | P04-03550 |
| Epidermal growth factor (EGF) from murine submaxillary gland | Sigma-Aldrich | E1257 |
| Foetal bovine serum | PAN Biotech | P30-3031 |
| Ham’s F12 medium, with 1.176 g/L NaHCO3 and l-glutamine | PAN Biotech | P04-14500 |
| Hydrocortisone | Sigma-Aldrich | H0888 |
| Insulin | Sigma-Aldrich | I6634 |
| Mitomycin C from Streptomyces caespitosus | Sigma-Aldrich | M4287 |
| Penicillin / Streptomycin (containing 10,000 U/ml Penicillin and 10 mg/ml Streptomycin) | PAN Biotech | P06-07100 |
| Trypsin 0.05% / ethylene-diamine tetra acetic acid (EDTA) 0.02% in PBS | PAN Biotech | P10-0235SP |
| LipofectamineTM RNAiMax | Thermo Fisher | 13778150 |
| FuGENE® HD | Promega | E2311 |
| Molecular biology | ||
| Kapa HiFi HotStart ReadyMix | Roche | KK2601 |
| Bbs I | New England Biolabs | R0539S |
| NEBuilder HiFi DNA assembly cloning kit | New England Biolabs | EE5520 |
| Monarch Total RNA Miniprep Kit | New England Biolabs | T2010S |
| LunaScript® Reverse Transcriptase (RT) SuperMix | New England Biolabs | E3010S |
| PowerUp™ SYBR™ Green Master Mix for qPCR | Thermo Fisher | A25742 |
| Absolute Q™ DNA Digital PCR Master Mix (5X) | Thermo Fisher | A52490 |
| Software | ||
| GraphPad Prism 10 | GraphPad Software LLC | |
| FACSDiva | BD Biosciences | |
| R-Studio | Posit (open source) | |
| R | R-project.org (open source) | |
| Cell Ranger | 10X Genomics | |
| Other | ||