KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-HA(Mouse) | BioLegend | Cat. #901515; RRID:AB_2565334 |
| Anti-HA(Rabbit) | BioLegend | Cat. #923502; RRID:AB_2565438 |
| Anti-FLAG(Mouse) | Sigma | Cat. #F1804; RRID:AB_262044 |
| Anti-FLAG(Rabbit) | Sigma | Cat. #F7425; RRID:AB_439687 |
| Anti-Phospho-ERK | Cell Signaling Technology (CST) | Cat.#9101; RRID:AB_331646 |
| Anti-ERK | CST | Cat. #9102; RRID:AB_330744 |
| Anti-Phospho-AKT(S473) | CST | Cat. #9271; RRID:AB_329825 |
| Anti-Phospho-AKT(T308) | CST | Cat. #9275; RRID:AB_329828 |
| Anti-AKT | CST | Cat. #9272; RRID:AB_329827 |
| Anti-MYC | Millipore-Sigma | Cat. #05–724; RRID:AB_309938 |
| Anti-Cleaved Caspase-3 | CST | Cat. #9661; RRID:AB_2341188 |
| Anti-CRAF | BD Biosciences | Cat. #610151; RRID:AB_397552 |
| Anti-BRAF | Santa Cruz | Cat. #sc-9002; RRID:AB_2067494 |
| Anti-GST | Santa Cruz | Cat. #sc-459; RRID:AB_631586 |
| Anti- Vinculin | Santa Cruz | Cat. #sc-73614; RRID:AB_1131294 |
| Anti-Ki-67 | Abcam | Cat. #16667; RRID:AB_302459 |
| Anti-GFP | CST | Cat. #2956; RRID:AB_1196615 |
| Anti-V5 | ThermoFisher | Cat. #MA5–15253; RRID:AB_10977225 |
| Cell Conditioning Solution (CC2) | Roche Diagnostics | Cat. #980–223 |
| DAPI | Akoya Biosciences | Cat. #FP1490 |
| Doxycycline (DOX) | Sigma | Cat. #D9891–100G |
| GTPγS | Sigma | Cat. #G8634 |
| 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) | Fisher | Cat. #M6494 |
| Opal Polaris 480 | Akoya Biosciences | Cat. #FP1500001KT |
| Opal 520 | Akoya Biosciences | Cat. #FP1487001KT |
| Opal 620 | Akoya Biosciences | Cat. #FP1495001KT |
| ProLong™ Gold Antifade Reagent | ThermoFisher | Cat. #P36934 |
| Bacterial and virus strains | ||
| E. coli BL21(DE3) | ThermoFisher | Cat. #694504 |
| S. cerevisiae EBY100 | Invitrogen | Discontinued |
| Biological samples | ||
| Patient Derived Xenografts (PDXs) | Ramsay Camp’s Lab, MUSC | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| His-Avi-TEV-HRAS (1–166) | Koide Lab, NYUSoM | N/A |
| His-Avi-TEV-HRAS (1–174) | Koide Lab, NYUSoM | N/A |
| His-Avi-TEV-KRAS (1–174) | Koide Lab, NYUSoM | N/A |
| His-Avi-TEV-NRAS (1–174) | Koide Lab, NYUSoM | N/A |
| His-Avi-TEV-KRAS(G12C) (1–174) | Koide Lab, NYUSoM | N/A |
| His-Avi-TEV-KRAS(G12D) (1–174) | Koide Lab, NYUSoM | N/A |
| His-Avi-TEV-KRAS(G12V) (1–174) | Koide Lab, NYUSoM | N/A |
| His-Avi-TEV-KRAS(K16N) (1–174) | Koide Lab, NYUSoM | N/A |
| His-Avi-TEV-KRAS(Q61L) (1–174) | Koide Lab, NYUSoM | N/A |
| GST-RAF-RBD | O’Bryan Lab, MUSC | N/A |
| Critical commercial assays | ||
| Gibson Assembly Master Mix | NEB | Cat. #E2611 |
| CellTiter-Glo Luminescent Cell Viability Assay | Promega | Cat. #G7571 |
| CloneAmp™ HiFi PCR Premix | Clontech | Cat. #639298 |
| Experimental models: Cell lines | ||
| HEK-293 | O’Bryan Lab, MUSC | N/A |
| HEK-293T | O’Bryan Lab, MUSC | N/A |
| COS-1 | O’Bryan Lab, MUSC | N/A |
| NIH/3T3 | O’Bryan Lab, MUSC | N/A |
| RASless MEFs | NCI-RAS Initiative | N/A |
| NCI-H1915 | ATCC | Cat. #CRL-5904 |
| LS1034 | NCI-RAS Initiative | N/A |
| HuP-T3 | Channing Der Lab, UNC, NC | N/A |
| PANC-1 | Gregory Thatcher Lab, UIC, IL | N/A |
| CF-PAC-1 | Channing Der Lab, UNC, NC | N/A |
| PSN-1 | Channing Der Lab, UNC, NC | N/A |
| HCT 116 | NCI-RAS Initiative | N/A |
| Hec1-A | Todd Waldman Lab, Georgetown University, Washington, DC | N/A |
| NCI-H1299 | Robert Winn Lab, UIC, IL | N/A |
| NCI-H1792 | Robert Winn Lab, UIC, IL | |
| NCI-H1944 | NCI-RAS Initiative | N/A |
| A375 | Andrew Aplin Lab, Thomas Jefferson University, PA | N/A |
| Experimental models: Organisms/strains | ||
| Athymic NU/NU nude mice | Charles River | Cat. #CR1 athymic nude mice |
| Athymic NU/NU nude mice | Taconic | Cat. #NCRNU |
| NSG (NOD-SCID gamma) mice | Jackson Laboratory | Cat. #005557 |
| Oligonucleotides | ||
| Monobody Gibson Forward Primer 5’→3’ (GACGATGACGACAAGGGATCCGTTTCTTCTGTTCC) | Eurofins | Custom |
| Monobody Gibson Reverse Primer 5’→3’ (TCAGTTATCTAGATCCGGTGGATCCCTAGGTACGGTAGTTAATCGAGATTGG) | Eurofins | Custom |
| KRAS Gibson Forward Primer 5’→3’ GGAGGACCTTCTAGCGGATCCATGACTGAATATAAACTTGTGGTAGTTGGAGCT | Eurofins | Custom |
| KRAS Gibson Reverse Primer 5’→3’ TCACCCTGAAGTTCTCAGGATCCTTACATAATTACACACTTTGTCTTTGAC | Eurofins | Custom |
| HRAS Gibson Forward Primer 5’→3’ GGAGGACCTTCTAGCGGATCCATGACAGAATAC | Eurofins | Custom |
| HRAS Gibson Reverse Primer 5’→3’ TCACCCTGAAGTTCTCAGGATCCTCAGGAGAGCAC | Eurofins | Custom |
| NRAS Gibson Forward Primer 5’→3’ GGAGGACCTTCTAGCGGATCCATGACTGAGTACAAACTGGTGGTG | Eurofins | Custom |
| NRAS Gibson Reverse Primer 5’→3’ TCACCCTGAAGTTCTCAGGATCCTTACATCACCACACATGGCAATCCC | Eurofins | Custom |
| Recombinant DNA | ||
| pCGN-HA tagged human KRAS WT | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human KRAS (G12V) | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human KRAS (G13D) | This Paper | N/A |
| pCGN-HA tagged human KRAS (G12C) | This Paper | N/A |
| pCGN-HA tagged human KRAS (G12D) | This Paper | N/A |
| pCGN-HA tagged human KRAS (G12R) | This Paper | N/A |
| pCGN-HA tagged human KRAS (G12S) | This Paper | N/A |
| pCGN-HA tagged human KRAS (G15A) | This Paper | N/A |
| pCGN-HA tagged human KRAS (Q61L) | This Paper | N/A |
| pCGN-HA tagged human KRAS (Q61H) | This Paper | N/A |
| pCGN-HA tagged human KRAS (Q61R) | This Paper | N/A |
| pCGN-HA tagged human KRAS (A146T) | This Paper | N/A |
| pCGN-HA tagged human KRAS [G12D(D69N)] | This Paper | N/A |
| pCGN-HA tagged human KRAS [G12V(D69N)] | This Paper | N/A |
| pCGN-HA tagged human KRAS [Q61L(D69N)] | This Paper | N/A |
| pCGN-HA tagged human KRAS [G12C(A59G)] | This Paper | N/A |
| pCGN-HA tagged human KRAS [G12D(A59G)] | This Paper | N/A |
| pCGN-HA tagged human KRAS [G12V(A59G)] | This Paper | N/A |
| pCGN-HA tagged human KRAS [Q61L(A59G)] | This Paper | N/A |
| pCGN-HA tagged human HRAS WT | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human HRAS(G12V) | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human HRAS(Q61L) | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human HRAS(K16N) | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human HRAS(D119N) | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human HRAS [G12V(D69N)] | This Paper | N/A |
| pCGN-HA tagged human HRAS [Q61L(D69N)] | This Paper | N/A |
| pCGN-HA tagged human NRAS(G12V) | This Paper | N/A |
| pCGN-HA tagged human NRAS(G12D) | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human NRAS(G13D) | O’Bryan Lab, MUSC | N/A |
| pCGN-HA tagged human NRAS(Q61L) | This Paper | N/A |
| pCGN-HA tagged human MEK(DD) | O’Bryan Lab, MUSC | N/A |
| pBabe MYC tagged human BRAF(V600E) | O’Bryan Lab, MUSC | N/A |
| pECFP | O’Bryan Lab, MUSC | N/A |
| pECFP-FLAG tagged NS1 | O’Bryan Lab, MUSC | N/A |
| pECFP-FLAG tagged R15 | This Paper | N/A |
| pECFP-FLAG tagged R15m10 | This Paper | N/A |
| pEYFP-HRAS WT | O’Bryan Lab, MUSC | N/A |
| pEYFP-HRAS (Q61L) | O’Bryan Lab, MUSC | N/A |
| pEYFP-NRAS (G12D) | O’Bryan Lab, MUSC | N/A |
| pCW57.1-CFP-FLAG tagged NS1 | This Paper | N/A |
| pCW57.1-CFP-FLAG tagged R15m10 | This Paper | N/A |
| pMD2.G VSV-G | Dider Trono Lab, | N/A |
| pCMVdR8.74 | Dider Trono Lab, University of Geneva | N/A |
| Software and algorithms | ||
| Prism | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| NIH-ImageJ | NIH | https://imagej.nih.gov/ij/ |
| Image Studio Lite | LICOR Biosciences | https://www.licor.com/bio/image-studio-lite/ |
| inForm® | Akoya Biosciences | https://www.akoyabio.com/phenoptics/software/inform-tissue-finder/ |
| Vectra Polaris | Akoya Biosciences | https://www.akoyabio.com/phenoptics/mantra-vectra-instruments/vectra-polaris/ |
| Other | ||
| Monobody sequence, R15: VSSVPTKLEVVAATPTSLLISWDASSSSVSYYRITYG ETGGNSPVQEFTVPGYYSTATISGLKPGVDYTITVYAYWYGYWSYISPISINYRT | Koide Lab, NYUSoM | N/A |
| Monobody sequence, R15m10: VSSVPTKLEVVAATPTSLLISWDASSSSVSYYRIT YGETGGNSPVQEFTVPGYYSTATISGLKPGVDYTITVYAVWQGVWRYVSPISINYRT | Koide Lab, NYUSoM | N/A |