KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-FLAG | Sigma-Aldrich | Cat # F1804 |
| Anti-TNRC6A | Bethyl Laboratories Inc. | Cat # A302–329A |
| Mouse monoclonal anti-α tubulin | Cell Signaling Technology | Cat # 3873T |
| Goat anti-mouse | LI-COR | Cat # 925–33210 |
| Goat anti-rabbit | LI-COR | Cat # 925–33211 |
| Anti-FLAG M2 beads | Sigma-Aldrich | Cat # A2220 |
| Bacterial and Virus Strains | ||
| DH5α competent cells | Invitrogen | Cat # 18258012 |
| One Shot MACH I™ TI | Invitrogen | Cat # 1493679 |
| BL21(DE3) Rosetta2 | Novagen | Cat # 71397–3 |
| DH10bac Mar Efficiency | Invitrogen | Cat # 1492020 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Prime STAR Max premix(2x) | Takara | Cat # R045 |
| Rapid DNA ligation kit | Roche | Cat # 11635379001 |
| Gibson assembly master mix | NEB | Cat # E2611S |
| Takara Z-taq™ | Takara | Cat # R006A |
| CellFectin® II reagent | Invitrogen | Cat # 10362–100 |
| TransIt-X2®system | Mirus | Cat # MIR 6006 |
| DpnI | NEB | Cat # R01768 |
| EcoRI | NEB | Cat # R0101T |
| EcoRV | NEB | Cat # R0195T |
| NotI | NEB | Cat # 0189S |
| SigmaFAST™ Protease inhibitor cocktail tablets, EDTA-free | Sigma-Aldrich | Cat # S8830–20TAB |
| T4PNK | Thermo Scientific | Cat # EK0031 |
| 4–12% gradient Bis-Tris gels | Life Technologies | Cat # NW04125BOX |
| γ−32P-ATP | Parkin-Elmer | Cat # BLU002A250UC |
| α−32P-GTP | Parkin-Elmer | Cat # BLU006H250UC |
| Phosphatase, alkaline (AP) | Roche | Cat # 11097975001 |
| Thermo Scientific RiboLock RNase Inhibitor | Thermo Scientific | Cat # E00381 |
| 3X FLAG peptides | Sigma-Aldrich | Cat # F4799 |
| Grace’s Insect Medium (1x) | Gibco, Life Technologies | Cat # 11605–094 |
| Penicillin Streptomycin | Gibco, Life Technologies | Cat # 15070–063 |
| DMEM (Dulbecco’s Modified Eagle Medium) | Gibco, Life Technologies | Cat # 11965–092 |
| FBS (Fetal Bovine Serum, Qualified) | Gibco, Life Technologies | Cat # 10437–028 |
| Opti-MEM® I | Gibco, Life Technologies | Cat # 31985–070 |
| 0.25% Trypsin-EDTA (1x) | Gibco, Life Technologies | Cat # 25200–056 |
| ESF 921 | Expression Systems | Cat # 96-001-01 |
| Deposited Data | ||
| Human AGO4 Structural data | This paper | PDB: 6OON |
| Human AGO1 (4KXT) | Nakanishi et al., 2013 | PDB: 4KXT |
| Human AGO1 (4KRE) | Faehnle et al., 2013 | PDB: 4KRE |
| Human AGO1 (4KRF) | Faehnle et al., 2013 | PDB: 4KRF |
| Human AGO2 (4F3T) | Elkayam et al., 2012 | PDB: 4F3T |
| Human AGO2 (4OLA) | Schirle et al., 2012 | PDB: 4OLA |
| Human AGO2 (4OLB) | Schirle et al., 2012 | PDB: 4OLB |
| Human AGO2 (4Z4D) | Schirle et al., 2015 | PDB: 4Z4D |
| Human AGO2 (6CBD) | Sheu-Gruttadauria et al., 2018 | PDB: 6CBD |
| AGO2 MID domain (3LUC) | Frank et al., 2010 | PDB: 3LUC |
| NcQDE2 (2YHA) | Boland et al., 2011 | PDB: 2YHA |
| Silkworm PIWI-clade Argonaute Siwi (5GUH) | Matsumoto et al., 2016 | PDB: 5GUH |
| Experimental Models: Cell Lines | ||
| Sf9 | Expression System | 94–001F |
| Tni | Expression System | 94–002F |
| HEK293T | ATCC | CRL-1573 |
| Oligonucleotides | ||
| AGO4WT-FW-GCGCGCCCATGGAGGCGCTGGGACC | This paper | N/A |
| AGO4WT-RV-GCGCGCGTCGACTCAGGCAAA ATAC | This paper | N/A |
| AGO2Y529E-FW- CGGCAAGACGCCCGTGGAAGCCGAG | This paper | N/A |
| AGO2Y529E-RV- ACGCGCTTGACCTCGGCTTCCACGGGC | This paper | N/A |
| AGO2E637A-FW- GCAGCAGCACCGGCAGGCGATCATACAAG |
This paper | N/A |
| AGO2E637A-RV -GGCGGCCAGGTCTTGTATGATCGCCTGCCGG | This paper | N/A |
| miR-20a-5p UAAAGUGCUUAUAGUGCAGGUAG | Dharmacon | N/A |
| miR-20a-3p ACCUGCACUAUAAGCACUUUAAG | Dharmacon | N/A |
| 60-nt miR-20a target RNA GGGAGAAACAAAAAUACCUACCUGCACUAUAAGCACUUUACCAUCUCAAACUUACUCAGA | Park et al., 2017b | N/A |
| 60-nt miR-20a mismatch target RNA GGGAGAAACAAAAAUACCUACCUGCACUAAUAGCACUUUACCAUCUCAAACUUACUCAGA | Dayeh et al., 2018 | N/A |
| Recombinant DNA | ||
| pFB-HTB | Invitrogen | Cat # 10584–027 |
| pCAGEN | Addgene | Cat # 11160 |
| SUMO fused pRSFDuet™−1 | Dayeh et al., 2018 | N/A |
| Software and Algorithms | ||
| iMOSFLM | https://www.mrc-lmb.cam.ac.uk/harry/imosflm/ | N/A |
| PHASER | McCoy et al., 2007 | N/A |
| PHENIX | www.phenix-online.org/ | N/A |
| PyMol | https://pymol.org/ | N/A |
| Coot | https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/ | N/A |
| Chimera | https://www.cgl.ucsf.edu/chimera/ | N/A |
| imageQuant | https://www.gelifesciences.com/en/us/shop/protein…/imagequant-tl-8-1-p-00110 | N/A |
| Image studio | https://www.licor.com/bio/image-studio/ | N/A |
| Prism | https://www.graphpad.com/scientific-software/prism/ | N/A |
| imageJ | https://imagej.nih.gov | N/A |