KEY RESOURCES TABLE

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Chemicals, Peptides, and Recombinant Proteins
ATP-γ-S, Lithium salt	Roche	Cat.# 11162306001
ATP, [γ−32P]	Perkin Elmer	Cat.# NEG002A250U
Critical Commercial Assays
Dual-Luciferase Reporter Assay system	Promega	Cat.# E1980
QuikChange II XL Site-Directed Mutagenesis Kit	Agilent	Cat.# 200522
Deposited Data
Raw Imaging Data	This Paper	http://dx.doi.org/10.17632/7psjn7wrfp.1
Experimental Models: Cell Lines
HEK293T Cell Line	Dr. Marcotrigiano	Devarkar et al., PNAS 2016
Oligonucleotides
5′ppp ss27 RNA 5′-pppAUACGUCCUGAUAGUUAGUAUCCAUCG-3′	Biosynthesis	N/A
5′OH ss27 RNA 5′-AUACGUCCUGAUAGUUAGUAUCCAUCG-3′	Dharmacon	N/A
5′OH ss25 RNA (for 3′ovg) 5′-ACGUCCUGAUAGUUAGUAUCCAUCG-3′	Dharmacon	N/A
complementary ss27 RNA 5′Bi-CGAUGGAUACUAACUAUCAGGACGUAU-3′ DY547	Dharmacon	N/A
5′ppp ss27 methylated RNA 5′-pppAUACGUCCUGAUAGUmUmAmGmUmAUCCAUCG-3′	Biosynthesis	N/A
complementary ss27 methylated RNA 5′Bi-CGAUGGAmUmAmCmUmAACUAUCAGGACGUAU-3′ DY547	Dharmacon	N/A
ss26 RNA chi (for stem dsRNA) 5′-dAdTdAdCdGUCCUGAUAGUUAGUAUCdCdAdTdC-3′	Dharmacon	N/A
complementary ss26 RNA chi intCy3 (for stem dsRNA) 5′-dCdGdAdTdGGAUACUAAC(Cy3)UAUCAGGAdCdGdTdA-3′	T riLink	N/A
complementary ss30 RNA chi (intCy3 at 13/17/20/24 nt from 3′ end) (for stopped-flow translocation assays) 5′Bi-dGdCdTCGAUGGAUACUAACUAUCAGGACGUAU-3′	T riLink	N/A
Recombinant DNA
pET28b 6xHis-SUMO-RIG-I	Jiang, et al. 2011	N/A
pET28b 6xHis-SUMO-HelRD	Jiang, et al. 2011	N/A
pET28b 6xHis SUMO-T667/671E RIG-I	This paper	N/A
pET28b 6xHis-SUMO-Δ673–685	Devarkar, et al. 2016	N/A
pET28b 6xHis-SUMO-C268F	This paper	N/A
pET28b 6xH is-SUMO-E373A	This paper	N/A
pcDNA 3.1 wt RIG-I	Devarkar, et al. 2016	N/A
pcDNA 3.1 empty vector	Devarkar, et al. 2016	N/A
pcDNA 3.1 E373A RIG-I	This paper	N/A
pcDNA 3.1 C268F RIG-I	This paper	N/A
pRL-TK (Renilla luciferase plasmid)	Devarkar, et al. 2016	N/A
pLuc125 (Firefly luciferase plasmid)	Devarkar, et al. 2016	N/A
Software and Algorithms
Kintek Explorer Enzyme Kinetics Software	Kintek	https://www.kintekexplorer.com
Other
TLC PEI Cellulose F	Millipore	Cat.# 105579