KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Bacterial and Virus Strains | ||
| Top10 Chemically Competent Cell | TransGen Biotech | Cat#CD101-01 |
| Rosetta (DE3) Chemically Competent Cell | TransGen Biotech | Cat#CD801-01 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| ULP1 | Homemade | N/A |
| T7 RNA polymerase | Homemade | N/A |
| MES | Sigma-Aldrich | Cat#M8250-250G |
| PEG 3000 | Sigma-Aldrich | Cat#81227-1KG |
| ethanol | Sigma-Aldrich | Cat#459836-1L |
| CHES | Sigma-Aldrich | Cat#C2885-100G |
| Sodium citrate | Hampton | Cat#HR2-235 |
| PEG 400 | QIAGEN | Cat#133086 |
| PEG 200 | Hampton | Cat#HR2-601 |
| 30% acrylamide and bis-acrylamide solution, 29:1 | LABLEAD | Cat#A3291-500ML |
| Critical Commercial Assays | ||
| JCSG Core Suite I | QIAGEN | Cat#130724 |
| JCSG Core Suite II | QIAGEN | Cat#130725 |
| JCSG Core Suite III | QIAGEN | Cat#130726 |
| JCSG Core Suite IV | QIAGEN | Cat#130727 |
| Crystal Screen | Hampton | Cat#HR2-110/112 |
| Index | Hampton | Cat#HR2-144 |
| Deposited Data | ||
| Structure of Apo_AcrIIC2 | This paper | PDB: 6J9K |
| Structure of NmeBH+AcrIIC2 | This paper | PDB: 6J9M |
| Structure of FnoBH+AcrIIC2 | This paper | PDB: 6J9L |
| Structure of NmeHNH+AcrIIC3 | This paper | PDB: 6J9N |
| Recombinant DNA | ||
| pRSF-Duet-1-His6-SUMO-NmeCas9 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas9_HNH508-667 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-455 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-241 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas9249-455 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas9Δ51-455 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas9ΔHNH | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-123 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W T54/D56A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W R62A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W R63A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W R66A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W R69A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W R70A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W R73A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W R74A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-NmeCas951-96W R69/R70/R73/R74A | This study | N/A |
| pRSF-Duet-1-His6-SUMO-SpyCas9 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-SpyCas951-96W | This study | N/A |
| pRSF-Duet-1-His6-SUMO-SpyCas9Nme-HNH | This study | N/A |
| pRSF-Duet-1-His6-SUMO-SaCas9 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-SaCas936-81W | This study | N/A |
| pRSF-Duet-1-His6-SUMO-SaCas9 Nme-HNH | This study | N/A |
| pRSF-Duet-1-His6-SUMO-CjeCas9 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-CjeCas937-82W | This study | N/A |
| pRSF-Duet-1-His6-SUMO-CjeCas9 Nme-HNH | This study | N/A |
| pRSF-Duet-1-His6-SUMO-FnoCas9 | This study | N/A |
| pRSF-Duet-1-His6-SUMO-FnoCas945-90W | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC1 | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC2 | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC2L36D | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC2Y11/I15/R20A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC2E18A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC2N23/D24/E25A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC2D42/D43A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC2Δ109-C | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC2K4/K10/R65/R66A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC3 | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC3R5A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC3R33A/D38A/Y40A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC3N60A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC3E92A/R103A | This study | N/A |
| pRSF-Duet-1-His6-SUMO- AcrIIC3M1S/F3S/L58S | This study | N/A |
| pET-28a-His6-MBP- NmeCas9 | This study | N/A |
| pET-28a-His6-MBP- NmeCas9Δ51-455 | This study | N/A |
| pET-28a-His6-MBP- AcrIIC2 | This study | N/A |
| pGEX-6P-1- AcrIIC3 | This study | N/A |
| Software and Algorithms | ||
| HKL2000 | Otwinowski and Minor, 1997 | http://www.hkl-xray.com/ |
| Phenix | Adams et al., 2010 | https://www.phenix-online.org/ |
| Coot | Emsley et al., 2010 | http://www2.mrc-lmb.cam.ac.uk/Personal/pemsley/coot/ |
| Pymol | The PyMOL Molecular Graphics System, Version 2.0 Schrödinger, LLC. | https://pymol.org/ |
| OriginPro 8 | OriginPro software | N/A |
| ImageJ | ImageJ software | N/A |
| Graphpad Prism | GraphPad software | N/A |
| Other | ||
| Amicon concentrators (3K) | Millipore | Cat#UFC900324 |
| Amicon concentrators (30K) | Millipore | Cat#UFC90302 |
| Ni Sepharose™ 6 FF beads | GE Healthcare | Cat#17-5318-03 |
| Glutathione Sepharose™ 4 Fast Flow beads | GE Healthcare | Cat#17-5132-02 |
| HiLoad™ 16/600 Superdex™ 200 pg | GE Healthcare | Cat#28-9893-35 |
| Superdex™ 75 10/300 GL | GE Healthcare | Cat#17-5174-01 |
| Superdex 200 10/300 GL | GE Healthcare | Cat#17-5175-01 |
| HiTrap™ Heparin HP (5 mL) | GE Healthcare | Cat#17-0407-01 |
| HiTrap™ SP Sepharose FF (5 mL) | GE Healthcare | Cat#17-5157-01 |