Reagent/Resource	Reference or Source	Identifier or Catalog Number
Experimental Models
HaCaT	Gift from Leonie Unterholzner (Lancaster University, UK)	RRID:CVCL_0038
HEK293	Gift from Caetano Reis e Sousa (The Francis Crick Institute, UK)	RRID:CVCL_0045
HEK293‐ISRE‐Fluc	Generated previously in our laboratory (Bridgeman et al, 2015)	N/A
HEK293T	Gift from Caetano Reis e Sousa (The Francis Crick Institute, UK)	RRID:CVCL_0063
MeWo	Gift from Graham Ogg (University of Oxford, UK)	RRID:CVCL_0445
THP1 cGAS KO	This study	N/A
THP1 Dual IFNAR2‐KO	Invivogen	Cat. #thpd‐koifnar2
THP1 Dual MyD88‐KO	Invivogen	Cat. #thpd‐komyd
THP1 Dual WT	Invivogen	Cat. #thpd‐nfis
THP1 IRF3 KO	This study	N/A
THP1 MAVS KO	This study	N/A
THP1 STING KO	Gift from Soren Paludan (Aarhus University, Denmark) (Holm et al, 2016)	N/A
THP1 TBK1 KO	Gift from Soren Paludan (Aarhus University, Denmark) (Holm et al, 2016)	N/A
VZV rOka	Gift from Jeffrey Cohen (NIH, Bethesda, USA) (Cohen & Seidel, 1993)	N/A
VZV‐ORF9ecDHFR	Gift from Paul R. Kinchington (University of Pittsburgh, USA) (Warner et al, 2021)	N/A
VZVORF9‐V5	Gift from Catherine Sadzot (University of Liege, Belgium) (Riva et al, 2013)	N/A
Recombinant DNA
hRIG‐I‐CARD	Gift from Andreas Pichlmair (Technical University Munich, Germany)	N/A
p125‐Luc	Gift from T. Fujita (Kyoto University, Japan) (Yoneyama et al, 2004)	N/A
p8.91	Gift from Greg Towers (University College London)	N/A
pcDNA/FLAG/hcGAS	This study	N/A
pcDNA/HA/hSTING	This study	N/A
pCMV‐3tag‐KSHV‐ORF52	Gift from Fanxiu Zhu (Florida State University, USA) (Wu et al, 2015)	N/A
pCMV‐VSV‐G	Gift from Bob Weinberg	Addgene_8454
pDONR/ORF22A	Gift from Jurgen Haas (University of Edinburgh, UK) (Uetz et al, 2006)	N/A
pDONR/ORF22B	Gift from Jurgen Haas (University of Edinburgh, UK) (Uetz et al, 2006)	N/A
pET‐6xHis‐SUMO2/hcGAS	Generated in previous study (Zhou et al, 2019)	N/A
pET‐6xHis‐SUMO2/ORF9	This study	N/A
pET‐6xHis‐SUMO2/ORF9‐N	This study	N/A
pET28a/FLAG‐GFP	This study	N/A
pET28a/FLAG‐ORF9	This study	N/A
pET28a/FLAG‐ORF9‐DM	This study	N/A
pGag‐eGFP	NIH AIDS reagent programme	#11468
pGEX6P1	Gift from Martin Reijns (University of Edinburgh, UK)	GE Healthcare
pGEX6P1/FLAG‐GFP	This study	N/A
pGEX6P1/FLAG‐ORF9	This study	N/A
pGEX6P1/hcGAS	Gift from Martin Reijns (University of Edinburgh, UK)	N/A
pGEX6P1/hcGAS‐HA	This study	N/A
pLenti/FLAG/GFP	This study	N/A
pLenti/FLAG/ORF9	This study	N/A
pLenti/FLAG/ORF9‐DM	This study	N/A
pLenti/FLAG/ORF9‐truncation‐I	This study	N/A
pLenti/FLAG/ORF9‐truncation‐II	This study	N/A
pLenti/FLAG/ORF9‐truncation‐III	This study	N/A
pLenti/FLAG/ORF9‐truncation‐IV	This study	N/A
pLenti/FLAG/ORF9‐truncation‐V	This study	N/A
pLenti/FLAG/ORF9‐truncation‐VI	This study	N/A
pLenti/FLAG/ORF9‐truncation‐VII	This study	N/A
pLenti/FLAG/ORF9‐truncation‐VIII	This study	N/A
pLenti6.3/V5/EMCV‐L	Generated in previous study (Hertzog et al, 2018)	N/A
pLenti6.3/V5/hcGAS	This study	N/A
pLenti6.3/V5/hSTING	This study	N/A
pLenti6.3/V5/VZV‐ORF	This study. Expression library for all VZV ORFs (see Figure EV2)	N/A
pRL‐TK	Promega	N/A
pX458/Ruby/hcGAS	This study	N/A
pX458/Ruby/hIRF3	This study	N/A
pX458/Ruby/hMAVS	Generated in previous study (Hertzog et al, 2018)	N/A
Antibodies
Donkey‐anti‐mouse‐HRP (WB, 1:2,000)	GE Healthcare	Cat. #NA931, RRID:AB_772210
Donkey‐anti‐rabbit‐HRP (WB, 1:2,000)	GE Healthcare	Cat. #NA934, RRID:AB_772206
Goat‐anti‐rabbit‐AlexaFluor647 (IF, 1:500)	Invitrogen	Cat. #A21246, RRID:AB_1500778
Mouse monoclonal beta‐actin‐HRP (WB, 1:10,000)	Sigma Aldrich	clone AC‐15, RRID:AB_262011
Mouse monoclonal FLAG (IP, 5 μg/sample)	Sigma Aldrich	clone M2, RRID:AB_262044
Mouse monoclonal FLAG‐HRP (WB, 1:10,000)	Sigma Aldrich	clone M2, RRID:AB_439702
Mouse monoclonal HA (IP, 5 μg/sample)	Invitrogen	clone 2‐2.2.14, RRID:AB_10978021
Mouse monoclonal RIG‐I (WB, 1:2,000)	Calteg Medsystems	clone Alme‐1, RRID:AB_2490189
Mouse monoclonal V5 (IP, 5 μg/sample)	Biolegend	Cat. #680602, RRID:AB_2566387
Mouse monoclonal V5‐FITC (IF, 1:400)	Invitrogen	Cat. #R963‐25, RRID:AB_2556567
Mouse monoclonal V5‐HRP (WB, 1:5,000)	Invitrogen	Cat. #R961‐25, RRID:AB_2556565
Mouse monoclonal VZV gE/GI (WB, 1:1,000)	GE Healthcare	Cat. #MAB8612, RRID:AB_2158042
Mouse monoclonal VZV gE/GI‐FITC (Flow cytometry, 1:500)	GE Healthcare, coupled in‐house to FITC	Cat. #MAB8612, RRID:AB_2158042
Mouse monoclonal VZV ORF62 (WB, 1:1,000)	Meridian Life Science	Cat. #C05107MA, RRID:AB_1772162
Rabbit monoclonal FLAG (IF, 1:800)	CellSignaling Technology	clone D6W5B, RRID:AB_2572291
Rabbit monoclonal hcGAS (WB, 1:1,000)	CellSignaling Technology	clone D1D3G, RRID:AB_2799712
Rabbit monoclonal hSTING (WB, 1:1,000)	CellSignaling Technology	clone D2P2F, RRID:AB_2732796
Rabbit monoclonal IRF3 (WB, 1:1,000)	CellSignaling Technology	clone D6I4C, RRID:AB_2722521
Rabbit monoclonal MyD88 (WB, 1:1,000)	CellSignaling Technology	clone D80F5, RRID:AB_10547882
Rabbit monoclonal pSTAT1 (Y701) (WB, 1:1,000)	CellSignaling Technology	clone 58D6, RRID:AB_561284
Rabbit monoclonal pSTAT2 (WB, 1:1,000)	CellSignaling Technology	clone D3P2P, RRID:AB_2800123
Rabbit monoclonal STAT1 (WB, 1:1,000)	CellSignaling Technology	clone 42H3, RRID:AB_2197984
Rabbit monoclonal STAT2 (WB, 1:1,000)	CellSignaling Technology	clone D9J7L, RRID:AB_2799824
Rabbit monoclonal TBK1 (WB, 1:1,000)	CellSignaling Technology	clone D1B4, RRID:AB_2255663
Rabbit polyclonal MAVS (WB, 1:1,000)	ENZO Life Science	Cat. #ALX‐210‐929‐C100, RRID:AB_2050916
Rabbit polyclonal VZV ORF9 (WB, 1:2,000)	Gift from Catherine Sadzot (University of Liege, Belgium) (Riva et al, 2013)	N/A
Oligonucleotides and sequence‐based reagents
VZV ORF library cloning PCR primers	This study	Appendix Table S1
Further cloning PCR primer	This study	Appendix Table S2
RT‐qPCR primers	This study	Appendix Table S3
VACV70mer F 5'‐ccatcagaaagaggtttaatatttttgtgagaccatcgaagagagaaagagataaaacttttttacgact‐3'	This study. Sequence described previously (Unterholzner et al, 2010)	N/A
VACV70mer R 5'‐agtcgtaaaaaagttttatctctttctctcttcgatggtctcacaaaaa‐tattaaacctctttctgatgg‐3'	This study. Sequence described previously (Unterholzner et al, 2010)	N/A
100 bp ISD F 5'‐ACATCTAGTACATGTCTAGTCAGTATCTAGTGATTATCTAGACATACATCTAGTACATGTCTAGTCAGTATCTAGTGATTATCTAGACATGGACTCATCC‐3'	This study. Sequence described previously (Zhou et al, 2021)	N/A
100 bp ISD R 5'‐GGATGAGTCCATGTCTAGATAATCACTAGATACTGACTAGACATGTACTAGATGTATGTCTAGATAATCACTAGATACTGACTAGACATGTACTAGATGT‐3'	This study. Sequence described previously (Zhou et al, 2021)	N/A
Chemicals, enzymes and other reagents
AlexaFluor‐488 (AF488) carboxylic acid (succinimidyl ester)	Thermo Fisher	Cat. #A20000
Antibody FITC Labeling Kit	abcam	Cat. #ab288100
BL21‐pLysS‐Rosetta	Novagene	Cat. #70954‐3
BL21‐RIL DE3 E. coli	Agilent	Cat. #230240
Blasticidin	gibco	Cat. #A1113903
CellTrace Yellow	Invitrogen	Cat. #C34567
DAPI	Invitrogen	Cat. #D1306
diABZi	Cambridge Bioscience	Cat #28054‐500 ug‐CAY
Dual Luciferase assay system	Promega	Cat. #E1910
Dynabeads Protein G	Invitrogen	Cat. #10004D
E. coli DNA	Invivogen	Cat. #tlrl‐ecdna
EZBlue Gel Staining Reagent	Sigma Aldrich	Cat. #G1041‐500ML
Gateway LR Clonase II Enzyme mix	Invitrogen	Cat. #11791020
Lipofectamine 2000	Invitrogen	Cat. #11668019
Lipofectamine LTX	Invitrogen	Cat. #15338100
LIVE/DEAD Fixable Violet Dead Cell Stain	Invitrogen	Cat. #L34955
Normal goat serum	Invitrogen	Cat. #10000C
oligo‐dT primers	Invitrogen	Cat. #18418012
PMA	Sigma Aldrich	Cat. #P1585‐1MG
Polybrene	Sigma Aldrich	Cat. #TR‐1003‐G
PreScission Protease	GE Healthcare	Cat. #GE27‐0843‐01
Protease Inhibitor Cocktail	CellSignaling Technology	Cat. #5871
Puyomycin	gibco	Cat. #A1113803
QuantiLuc substrate	Invivogen	Cat. #rep‐qlc1
SuperScript IV Reverse Transcriptase	Invitrogen	Cat. #18090050
SYBR GreenER master mix	Invitrogen	Cat. #11762100
TaqMan Universal PCR Master Mix	Invitrogen	Cat. #4304437
Trimethoprim	Sigma Aldrich	Cat. #46984‐250MG
α32P‐ATP	Perkin Elmer	Cat. #NEG003X250UC
Software
Web‐based iBright Image Analysis software	Thermo Fisher
FlowJo	FlowJo, LLC
SnapGene	GSL Biotech
ApE	M. Wayne Davis, The University of Utah
Fiji 2.0.0	Schindelin et al (2012)