Key resources table.
| Reagent type (species) or source |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (human) | HEK293T RIG-I KO | Zhu et al. (2014) | Growth in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) as monolayer |
|
| Strain, strain background (Escherichia coli) |
BL21 (DE3) Rosetta | Novagene | ||
| Strain, strain background (Escherichia coli) |
DH10multiBac | GenevaBiotech | ||
| Strain, strain background (Spodoptera frugipeda) |
Sf21 insect cells | Thermo Fisher Scientific |
11497013 | Growth in SF-900 III serum-free medium |
| Strain, strain background (Trichoplusia ni) |
High Five insect cells | Thermo Fisher Scientific |
B85502 | Growth in Express Five serum-free medium supplemented with 10 mM L-glutamine |
| Recombinant DNA reagent |
pcDNA5/FRT/TO | Thermo Fisher Scientific |
V652020 | |
| Recombinant DNA reagent |
pcDNA5/FRT/TO-FLAG/ HA-RIG-I and various mutants of the same construct |
Lässig et al. (2015)
and this paper |
Progenitors: PCR, DDX58 (cDNA) and pcDNA5/FRT/TO |
|
| Recombinant DNA reagent |
p-125luc | Yoneyama et al. (1996) | Firefly luciferase controlled by an interferon-β promoter |
|
| Recombinant DNA reagent |
pGL4.74 | Promega | E6921 | Constitutive expression of a Renilla luciferase |
| Recombinant DNA reagent |
pFBDM | Berger et al. (2004) | ||
| Recombinant DNA reagent |
pFBDM-His-RIG-I and various mutants of the same construct |
Lässig et al. (2015)
and this paper |
Progenitors: PCR, DDX58 (cDNA) and pFBDM |
|
| Recombinant DNA reagent |
pETM11-SUMO3GFP | EMBL Heidelberg, H. Besir |
https://www.embl.de/pepcore/pepcore_services/cloning/sumo/ | |
| Recombinant DNA reagent |
pETM11-SUMO3-RIG-I- Δ2CARD-C268F |
This paper | Progenitors: PCR, DDX58 (cDNA) and pETM11-SUMO3GFP |
|
| Sequence-based reagent |
19mer 5' triphosphate dsRNA |
InvivoGen | tlrl-3prna | 1 µg/mL, 5’-pppGCAUGC GACCUCUGUUUGA-3 |
| Sequence-based reagent |
14mer dsRNA | Dharmacon | 5'-CGACGCUAGCGUCG-3' | |
| Sequence-based reagent |
Cy3-hpRNA | Biomers | 5'-Cy3-CCACCCGCCCCCCUAGU GAGGGGGGCGGGCC-3' |
|
| Chemical compound, drug |
Lipofectamine 2000 | Thermo Fisher Scientific |
11668019 | Used at 2.5x excess compared to RNA/DNA mass |
| Chemical compound, drug |
MANT-ATP | Jena Bioscience | NU-202 | |
| Chemical compound, drug |
MANT-ATPγS | Jena Bioscience | NU-232 | |
| Chemical compound, drug |
[γ-32P]ATP | Hartmann Analytic | SRP-301 | 10 nM spiked with 3 mM unlabeled ATP |
| Commercial assay or kit |
Dual-Luciferase Reporter Assay System |
Promega | E1910 | |
| Software, algorithm |
XDS, XSCALE | Kabsch (2010) | http://xds.mpimf-heidelberg.mpg.de/ | |
| Software, algorithm |
PHASER |
McCoy et al. (2007); Winn et al. (2011) |
http://www.ccp4.ac.uk/ | |
| Software, algorithm |
Coot | Emsley et al. (2010) | https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/ | |
| Software, algorithm |
PHENIX | Afonine et al. (2012) | https://www.phenix-online.org/ | |
| Software, algorithm |
Pymol | Schrödinger | https://pymol.org/2/ | |
| Software, algorithm |
R | R Development Core Team (2013) | https://www.r-project.org/ |