KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Chemicals, Peptides, and Recombinant Proteins | ||
| Gibson Assembly Master mix | New England Biolabs | #E2611 |
| Phusion High-Fidelity DNA Polymerase | ThermoFisher Scientific | #F530 |
| DpnI-FD | ThermoFisher Scientific | #FD1703 |
| BsaI-HFv2 | New England Biolabs | #R3733 |
| Hifi-Assembly DNA Assembly | New England Biolabs | #E5520 |
| SYBR™ Gold | ThermoFisher Scientific | #S11494 |
| T7 Endonuclease 1 | New England Biolabs | #M0302L |
| DMEM (Medium for mammalian cell culture) | GIBCO | #11965092 |
| Fetal Bovine Serum (For mammalian cell culture) | Sigma Aldrich | #F4135 |
| Penicillin-Streptomycin (For mammalian cell culture) | Sigma Aldrich | #P4333 |
| PolyFect transfection reagent | QIAGEN | #3011 |
| High Fidelity 2X PCR Master Mix | New England Biolabs | #M0541S |
| DreamTaq Green PCR Master Mix 2X | ThermoFisher Scientific | #K1081 |
| DNA Gel Elution Buffer | New England Biolabs | #E7324A |
| SUPERase· In RNase Inhibitor | ThermoFisher Scientific | #AM2696 |
| Critical Commercial Assays | ||
| DNA Clean & Concentrator-5 | Zymo Research | #D4004 |
| NEBNext Small RNA Library Prep Set for Illumina | New England Biolabs | #E7330 |
| DNeasy Blood and Tissue Kit | QIAGEN | #69504 |
| Ni-NTA agarose resin | QIAGEN | #30210 |
| Superdex 200 10/300 | GE Healthcare | #28990944 |
| Experimental Models: Cell Lines | ||
| Neuro-2a | ATCC | CCL-131 |
| Human HEK293T | ATCC | ATCC CRL-3216 |
| HEK293T-TLR-MCV1 variant | This study | N/A |
| HEK293T-TLR-MCV1-AcrIIA5 | This study | N/A |
| Oligonucleotides | ||
| See Table S1 for sequences of sgRNAs used | This study | N/A |
| N66H70N73aA5F GAAATCCGTCTGTCCAATgcCAGTGCTGATgcTAAATACgcTGATCTTGAGAATGGGCG | Integrated DNA Technologies | N/A |
| N66H70N73aA5R CGCCCATTCTCAAGATCAgcGTATTTAgcATCAGCACTGTcATTGGACAGACGGATTTC | Integrated DNA Technologies | N/A |
| D50R62aA5F AAGTATGAAGGcTTCTGCCTATAAAGACTTTGGAAAATATGAAATCgcTCTGTCCAATCA | Integrated DNA Technologies | N/A |
| D50R62aA5R TGATTGGACAGAgcGATTTCATATTTTCCAAAGTCTTTATAGGCAGAAgCCTTCATACTT | Integrated DNA Technologies | N/A |
| D74K8588aA5F AAATACCATGcTCTTGAGAATGGGCGTCTTATCGTGAACATCgcAGCATCAgcGCTGAAC | Integrated DNA Technologies | N/A |
| D74K8588aA5R GTTCAGCgcTGATGCTgcGATGTTCACGATAAGACGCCCATTCTCAAGAgCATGGTATTT | Integrated DNA Technologies | N/A |
| Recombinant DNA | ||
| pGeoCas9-sgRNAMu | Harrington et al., 2017 | https://benchling.com/s/seq-IXSEQMlloSIZmGtpNsld/edit |
| pHpaCas9sgRNAMu | Lee et al., 2018 | N/A |
| pNme1Cas9sgRNAMu | Thavalingam et al., 2019 | N/A |
| pCjeCas9sgRNAMu | Thavalingam et al., 2019 | N/A |
| pBoeCas9sgRNAMu | This study | N/A |
| pKlaCas9sgRNAMu | This study | N/A |
| pCdiCas9sgRNAMu | This study | N/A |
| pSauCas9sgRNAMu | This study | N/A |
| pSpyCas9sgRNAMu | This study | N/A |
| pFnoCas9sgRNAMu | This study | N/A |
| pCDF-1b | Novagen | #71330-3 |
| pCDF-ACRE2 | Harrington et al., 2017 | N/A |
| pCDF-ACRIIC1Nme | Harrington et al., 2017 | N/A |
| pCDF-ACRIIC2Nme | Thavalingam et al., 2019 | N/A |
| pCDF-ACRIIC3Nme | This study | N/A |
| pCDF-ACRIIC4Hpa | Lee et al., 2018 | N/A |
| pCDF-ACRIIC5Smu | Lee et al., 2018 | N/A |
| pCDF-ACRIIA2Lmo | This study | N/A |
| pCDF-ACRIIA3Lmo | This study | N/A |
| pCDF-ACRIIA4Lmo | This study | N/A |
| pCDF-ACRIIA5SthD4276 | This study | N/A |
| pCDF-ACRIIA5SthD1126 | This study | N/A |
| pCDF-ACRIIA5Efa | This study | N/A |
| pCDF-ACRIIA5Dpi | This study | N/A |
| pCDF-ACRIIA5Lsa | This study | N/A |
| pCDF-ACRIIA5G572 | This study | N/A |
| pCRISPathBrick | Cress et al., 2015 | Addgene #65006 |
| pCM-str | J. Nodwell Lab | N/A |
| TOPO® vector | ThermoFisher Scientific | #K4500-01 |
| 6x-His-tagged Nme1Cas9-sgRNA in pMCSG7 | Pawluk et al., 2016 | N/A |
| 6x-His-tagged SpyCas9-sgRNA | This study | N/A |
| SpyCas9-sgRNA | Ran et al., 2013 | Addgene #62988 |
| SauCas9-sgRNA | Ran et al., 2015 | Addgene #61591 |
| pEJS24-pCSDest2-SpyCas9-NLS-3XHA-NLS | Bolukbasi et al., 2015 | Addgene #69220 |
| pEJS424-pCSDest2-NmeCas9-NLS-3XHA-NLS | Pawluk et al., 2016 | Addgene #87448 |
| pEJS485-pCSDest2-SauCas9 | S. Wolfe lab | N/A |
| pX404-CjeCas9 | F. Zhang lab | Addgene #68338 |
| pEJS433 pCSDest2-AcrIIC1 | Pawluk et al., 2016 | Addgene #85679 |
| p611-pCSDest2-AcrIIA4 | This study | N/A |
| pEJS1004-pCSDest2-AcrIIA5-NLS-FLAG | This study | N/A |
| pEJS1005-Lenti-mammalian c.o.AcrIIA5-FLAG-NLS-HygR | This study | N/A |
| Software and Algorithms | ||
| FigTree 1.4.3 | Institute of Evolutionary Biology, University of Edinburgh | http://tree.bio.ed.ac.uk/software/ |
| FlowJo® v10.4.1. | FlowJo LLC | N/A |
| Gen5 2.09 | BioTek Instruments Inc | https://www.biotek.com/products/software-robotics-software/gen5-microplate-reader-and-imager-software/software/ |