Unravel the Mysteries of STING:
A Hub for Innate Immune Responses to Nucleic Acids

 InvivoGen sas Europe (Toulouse, France)
Founded in 1977 by Pr. Gerard Tiraby

 InvivoGen Inc. America (San Diego, U.S.)
US subsidiary created in 1997

 InvivoGen Ltd. Asia (Hong Kong, China)
Asian subsidiary incorporated in 2014

Our bodies are constantly being exposed to infectious agents

Research in defense of infectious agents is lasting for centuries

Chinese began practicing a formof immunization by drying andinhaling powders derived fromthe crusts of smallpox lesions in1000 A.D.

Edward Jenner
developed the first
vaccine(Small pox
vaccine) in the worldin 1798.

Élie Metchnikoff
discovered phygocytes
in 1882, won the 1908
Nobel Prize inPhysiology or
MedicineIn 1989

Cited by 3442 articles

Innate Immune System
non specificfirst line of defenseimmediate responsememory is non-specific

Adaptive Immune Systemspecific
second line of defense
slow responselong term memory

Innate immune system active Adaptive Immune system: pathogen-associated molecularpatterns (PAMPs) are recognized by pattern recognition receptors (PRRs) , in turn trigger B7 (CD80/86)
signal and cytokinesJaneway, C. A. Jr. Approaching the asymptote? Evolution and revolution in immunology. Cold Spring Harb. Symp. Quant. Biol. 54, 1–13 (1989).

Innate immune system active Adaptive Immune system: pathogen-associated molecularpatterns (PAMPs) are recognized by pattern recognition receptors (PRRs) , in turn trigger B7 (CD80/86)
signal and cytokines

Janeway, C. A. Jr. Approaching the asymptote? Evolution and revolution in immunology. Cold Spring Harb. Symp. Quant. Biol. 54, 1–13 (1989).
Ruslan Medzhitov. et al. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature. 1997

Recognition of microbial pathogens: Microbial pathogens
expressed highly conserved structural motifs named pathogenassociated molecular patterns (PAMPs) which could be specificallyrecognized by pattern recognition receptors (PRRs).

PRRs responses to PAMPs on cell surface and in endosome

PRRs responses toreplicating microbes
in cytoplasm

STING research

Publication 711

Publication 849
Publication 1439

2008~April of 2020

Interferon
Stimulatory DNA
(ISD)

STING

TBK1

Product Box
Product
ISD Naked

Cat. code
tlrl-isdn

STING – Stimulator of interferon
genes (also know as TMEM173, ERIS,MITA and MPYS) is essential for
DNA sensing in cytosol, where inturn induce type I interferon.
However, cytosolic DNA does notdirectly bind with STING.

Sendai Virus

MITA

TBK1

IRF3

IRF3

IFNs

IFNs

Ishikawa H. et al., Nature. 2008
Ishikawa H. et al., Nature. 2009

Zhong B. et al., Immunity. 2008
Zhong B. et al., Immunity. 2009

Guijun Shang, et al. Nature. 2019

Product Box

DNA

cGAS

DAI

IFI16

IFIX

cyclic-di-GMP
cyclic-di-AMP(Second(Se
Messenger)
Mess

MRE11



TBK1
IRF3
IFNs

STING

DDX41

Product

Cat. code

SensorPoly(dA:dT)

tlrl-patn

DAI

Poly(dG:dC)

tlrl-pgcn

DAI

HSV-60tlrl-hsv60n IFI16

VACV-70tlrl-vav70n IFI16

ISD Naked

tlrl-isdn

controversial

G3-YSD [NEW] tlrl-ydna

cGAS

c-di-GMP

tlrl-cdg

DDX41, STING

c-di-AMP

tlrl-cda

DDX41, STING

c-di-IMP

tlrl-cdi

DDX41, STING

Ishikawa H. et al., Nature. 2008
Takaoka,A. et al.,Nature. 2007
Upton. et al., Cell Host Microbe. 2012
Unterholzner,l., et al., Nat Immnol. 2010
Diner B.A., et al., Mol syst Biol. 2015
Horan, K.A., et al., J. Immunol. 2013
Kondo,T., et al., PNAS. 2013
Zhang, Z.Q., et al., Nat Immnol. 2011
Burdette, D.L., et al., Nature. 2011
Sun L. et al., Science. 2013
Wu J. et al., Science. 2013

DNA
cGAS is the only
cytosolic DNA
sensor that triggers
IFN

cGAS

Elizabeth E. Gray. et al.,Immunity. 2016

cGAMP

TBK1

IRF3

IFNs

STING

A Narrowing Field of Cytosolic DNA Sensors Triggering IFN Production
Genetic data now point to cyclic-GMP-AMP synthase (cGAS) as the only viable IFN-inducing
DNA sensor, depicted in this illustration as the last man standing. It remains unclear
whether other putative cytosolic DNA sensors, previously proposed to induce type I IFN
expression in response to dsDNA, are truly dead and buried or whether these proteins will
have roles in response to specific stimuli in certain cellular contexts.

Russell E. Vance. Immunity.2016

6
IFI1

cG
AS

DNA

K.L. Jønsson et al. IFI16 is required for DNA sensing in human macrophages by promoting production and function ofcGAMP. Nature Communication.2017

6

IFI1

ST
IN
G

cGAMP

IFI16 is required for DNA
in HaCaT keratinocytes.

IFI16 is required for STING
response to cGAMP .

Interaction between endogenous IFI16 and cGAS
that was enhanced by stimulation with DNA

Jessica F. Almine et al. IFI16 and cGAS cooperate in the activation of STING during DNA sensing in human keratinocytes.
Nature Communication.2017

Product Box

DNA

cGAS

Product
Cat. code
3'3'-cGAMP tlrl-nacga
2'3'-cGAMP tlrl-nacga23
2’2'-cGAMP tlrl-nacga22

Cyclic GMP-AMP synthase(cGAS)
Sun L. et al., Science. 2013

cGAMP

Sensor
STING
STING
STING

Cyclic guanosine
monophosphate-adenosine
monophosphate(cGAMP)

Metazoan CDNs

2'3'-cGAMP

Wu J. et al., Science. 2013
Microbial CDNs

TBK1

IRF3

IFNs

STING
3'3'-cGAMPcGAS is not cytosolic sensor

Plasma membrane cGAS cannot be detected in the absence of benzonasecGAS Associates with the Plasma Membrane

Phosphoinositide Interactions Position cGAS at the Plasma Membrane to
Ensure Efficient Distinction between Self- and Viral DNA. Katherine C.
Barnett, Julia M. Coronas-Serna, Wen Zhou, Michael J. Ernandes, Anh
Cao,1 Philip J. Kranzusch and Jonathan C. Kagan. Cell. 2019

Mislocalized cGAS mutants drive interferon responses to genotoxic stressMolecular Mechanisms

STING

cGAS

2020

Infectious Diseases

I

C

Caner

A

Autoimmunity

cGAS
STING

A

Adjuvant

Infectious Diseases

I

cGAS
STING

 Invasion
 Evasion
 Interplay

Andrea Ablasser and Sun Hur. Nature Immunology. 2019

STING and Infectious Diseases - Invasion
cGAS/STING & Mycobacterium tuberculosis

cGAS Is Essential for Type I IFN Responses Triggered by Mtb

Product

5’ppp-dsRNA

Cat. code
tlrl-3prna-100

THP1-Dual™ KO-STING Cells

thpd-kocgas

THP1-Dual™ KO-cGAS Cells

thpd-kostg

cGAMP Shuttling Restores Type I IFN Responses
Ruth Wassermann, et. al. Mycobacterium tuberculosis Differentially
Activates cGAS- and Inflammasome-Dependent Intracellular Immune
Responses through ESX-1. Cell Host & Microbe. 2015

STING and Infectious Diseases - Invasion
STING & Mycobacterium tuberculosis
M. tuberculosis genome encodes a diadenylate cyclase enzyme (disA) thatsynthesizes c-di-AMP from ATP or ADP.

Bappaditya Dey
Dey,etet al.
al A bacterial cyclic dinucleotide
activates the cytosolic surveillance pathway and mediates
innate resistance to tuberculosis. Nature Medicine. 2015

Product
RAW-Lucia™ ISG Cells

Cat. coderawl-isg

RAW-Lucia™ ISG KO-STING Cells

rawl-kostg

STING
Infectious and 
 Diseases   – Invasion
STING & HIV

Sunnie M.Yoh, , et al. PQBP1 Is a Proximal Sensor of thecGAS-Dependent Innate Response to HIV-1 . Cell .2016

STING and Infectious Diseases – Evasion
STING & HIV

THP-1

Mitochondriallocalized NLR, NLRX1

BMDM

Guo et al., NLRX1 Sequesters STING to Negatively Regulate the Interferon Response,
Thereby Facilitating the Replication of HIV-1 and DNA Viruses. Cell Host & Microbe, 2016

STING and Infectious Diseases – Evasion
STING & HCMV-UL82 Inhibits STING-Mediated Signaling

Yu-Zhi Fu, et al. Human Cytomegalovirus Tegument
Protein UL82 Inhibits STING-Mediated Signaling to
Evade Antiviral Immunity. Cell Host & Microbe. 2017

HCMV

STING
ER Tracker

UL82

VectorMock-UL82 Impairs the Trafficking of STINGSTING and Infectious Diseases – Evasion
STING & Dengue
DengueProtease
B3
NS2



NS2B

cGAS

DegradehSTIIN
NG

mS

N
TI

G

[1] DNS2B3 Cleave
hSTING but not
mSTING

Cleave[2] NS2B degrades cGAS
[1] Yu, C.Y. et al. Dengue virus targets the adaptor protein MITA to subvert host innate
immunity. PLoS Pathog. 2012.
[2] Sebastian Aguirre. et al. Dengue virus NS2B protein targets cGAS for degradation andprevents mitochondrial DNA sensing during infection NATURE MICROBIOLOGY 2017.

Product

Cat. code

Poly(dA:dT)

tlrl-patn-1

VACV-70tlrl-vav70n

STING and Infectious Diseases – Evasion
STING & Poxvirus

Poxvirus

Poxin

cGAS




2’3’-cGAM
P- Vaccinia virus(VACV) degrades 2’3’-cGAMP- Poxin, a virus nuclease is identified
Product
Cat. code
3'3'-cGAMP tlrl-nacga
2'3'-cGAMP tlrl-nacga23
c-di-GMP
tlrl-cdg
c-di-AMP
tlrl-cda- Poxin specifically degrades 2’3’-cGAMPJames B. eaglesham. et al.Viral and metazoan poxins are cGAMP-specific nucleases that restrict
cGAS–STING signalling. Nature. 2019.

STING and Infectious Diseases – Evasion
STING & Mycobacterium tuberculosis
Time kinetics of 23-cGAMP cleavage by CdnP

Product
3'3'-cGAMP
2'3'-cGAMP
c-di-GMP
RAW-Blue ISG Cells
RAW-Blue ISG KO-STING Cells
THP-1-Dual Cells

Cat. code
tlrl-nacga
tlrl-nacga23
tlrl-nacdg
raw-isg
rawl-kostg
thpd-nfis

Ruchi Jain dey, et. al. Inhibition of innate immune cytosolic surveillance byan M. tuberculosis phosphodiesterase. Nature Chemical Biology. 2017

STING and Infectious Diseases – Evasion

James B Eaglesham and Philip J Kranzusch. Conserved strategies
for pathogen evasion of cGAS–STING immunity. Current Opinion in
Immunology. 2020

STING and Infectious Diseases – Evasion

Nicolas Vabret, et. al. Immunology of COVID-19: current state of the science. Immunity. 2020

STING and Infectious Diseases – Interplay
STING & TLR7 - Malaria-Mice deficient in cGAS, STING, MDA5, MAVS, but not in TLR7
signaling molecules, are protected from YM Infection-SOCS1 inhibit
Myd88dependentanti-malaria

Xiao Yu, et al. Cross-Regulation of Two Type I Interferon
Signaling Pathways in Plasmacytoid Dendritic Cells
Controls Anti-malaria Immunity and Host Mortality.
Immunity. 2016

-SOCS1 associatewith Myd88

STING and Infectious Diseases – Interplay
STING & Inflammasome – DNA Viurs-Inflammasome Defect Augments
Cytokine Production-Inflammasome Inhibition Potentiates
DNA Virus-Triggered Signaling-cGAS Is Recruited and Cleaved by Caspase-11and Caspase-1during
Inflammasome Activation

Simon Heidegger, et al. Cutting Edge in IFN Regulation:
Inflammatory Caspases Cleave cGAS. Immunity. 2017
Yutao Wang, et al. Inflammasome Activation Triggers
Caspase-1-Mediated Cleavage of cGAS to Regulate
Responses to DNA Virus Infection Immunity. 2017

Product Box

Products
HSV-60 Naked

Cat.
tlrl-hsv60n

VACV-70tlrl-vav70n

VX-765Inh-vx765i-1

STING and Infectious Diseases – Interplay
STING & Gasdermin D - Bacteria

Francisella novicida

Products
Ultrapure LPS E.coli O111:B4
2’3’-cGAMPPoly I:C
Pam3CSK4
Nigericin
Ionomycin

Cat.
tlrl-3pelps
tlrl-nacga23-1
tlrl-pic
tlrl-pms
tlrl-nig
Inh-ionIshita Banerjee, et al. Gasdermin D Restrains Type I Interferon Response to Cytosolic DNA by
Disrupting Ionic Homeostasis. Immunity. 2018
Rebecca Feltham and James E. Vince. Ion Man: GSDMD Punches Pores to Knock Out cGAS.
Immunity. 2018

Pyroptosis and Necroptosis Reporter Cell Lines
THP1-HMGB1-LuciaTM Cells
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STING and Infectious Diseases - Bacteria
STING mediates Autophagy

Product
c-di-AMP
Rapamycin
Bafilomycin AI
Wortmannin
HeLa-Difluo™ hLC3 Cells

Cat. code
tlrl-nacda
tlrl-rap
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tlrl-wtm
heldf-hlc3

Julien Moretti, et. al. STING Senses Microbial Viability to OrchestrateStress-Mediated Autophagy of the Endoplasmic Reticulum.Cell. 2017

STING and Autophagy"%%##% &  !

http://www.invivogen.com/autophagy      

C

cGAS
STING

Caner
Cold

Hot

Demaria et al, Nature 2019 (574)

STING and Cancer

2011

2014

STING and IRF3 Are Required for CD8+ T Cell Priming in vivo

STING pathway can play a critical rolein the therapeutic efficacy of cancer
immunotherapies.

Product
B16-Blue™ ISG Cells

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bb-ifnabg

STING and Cancer

DMXAA in anti-tumor therapy

Product Box
Product

DMXAACat. code

tlrl-dmx

B16.SIY tumor cells in WT C57BL/6 miceAnti-tumor

Immunologic Memory
Leticia Corrales, et al. Cell Reports. 2015

STING and Cancer

DMXAA = 5,6-dimethylxanthenone-4-acetic acid
STING sequence

DMXAA binding affinity

Ref.

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Stop in Phase III

DMXAA
293T-Dual™ hSTING-A162 Cells
THP1-Dual™ KI-hSTING-A162 Cells

Cat. code
tlrl-dmx
293d-a162
thpd-a162

STING and Cancer

New STING ligand in anti-tumor therapy
Screened by

THP1-Blue™ ISG Cells-ML RR-S2 CDA:
Mixed linkage dithio-(RP, RP)-[cyclic[A(2’,5’) pA(3’,5’)p-ADU-S100 (2’3’-c-di-AM(PS)2 (Rp,Rp)): Phase I/II

Product Box
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Cat. code
THP1-Blue™ ISG Cells
thp-isg
QUANTI-Blue™
Rep-qb1/2
2’3’-c-di-AM(PS)2 (Rp,Rp) tlrl-nacda2r
Leticia Corrales, et al. Cell Reports. 2015

Data for proving human STING specificity

STING and Cancer

New STING ligand in anti-tumor therapy
Screened by

THP1-Blue™ ISG Cells-ML RR-S2 CDA:
Mixed linkage dithio-(RP, RP)-[cyclic[A(2’,5’) pA(3’,5’)p-ADU-S100 (2’3’-c-di-AM(PS)2 (Rp,Rp)): Phase I/II

Product Box
Product
Cat. code
THP1-Blue™ ISG Cells
thp-isg
QUANTI-Blue™
Rep-qb1/2
2’3’-c-di-AM(PS)2 (Rp,Rp) tlrl-nacda2r
Leticia Corrales, et al. Cell Reports. 2015

The author could address simple STING
specific evidence through THP1-Dual KOSTING cells
Human cell line
Products
THP1-Dual™ cells
THP1-Dual™ KO-STING cellsCat.
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thpd-kostg

STING and Cancer

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STING and Cancer

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DMXAA
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2’3’-c-di-AM(PS)2 (Rp,Rp) tlrl-nacda2r

WT C57BL/6 Mice Lung

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STING and Cancer

ADU-S100

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STING and Cancer- Intravenous administration- A small molecule STING agonist that is not a cyclic dinucleotide

CT-26 Colon Tumor
Joshi M. ramanjulu, et al. Design of amidobenzimidazole STING receptor agonists with systemic activity. Nature. 2018

STING and Cancer

Novel delivery system
Auxotrophic E. coli Nissle (EcN)
engineered to produce high levels of
the STING agonist c-di-AMP

Synlogic Cooperate Presentation, 2019

Daniel S. Leventhal, et al.        
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STING and Cancer

Exosome

Su Chul, Jiang. Selective Activation of Antigen Presenting Cells by exoSTING Enhances Tumor Antigen-Specific Immune
Response. Presented at the 34thAnnual Meeting of the Society for Immuno-Therapy of Cancer, November 6.

STING and Cancer

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Patent application (WO2019129880) for an ADC with a proprietary CDN(to be commercialised in 2020)

Cathepsin sensitive self-immolating linker

Proprietary STING Agonist

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InvivoGen Proprietary CDN
Novel Cyclic Adenosine-Inosine Monophosphate Analogs

InvivoGen literature

Lioux et al., 2016, J. Med. Chem.
10.1021/acs.jmedchem.6b01300

CL592 and CL656 is InvivoGen Patented CDN
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cAIMP

cAIM(PS)2 Difluor (Rp/Sp)

2’3’-c-di-AM(PS)2 (Rp,Rp)

Synonym: 3’3’-cAIMP sodium salt,

Synonym: (Rp/Sp) c-[2’FdAM(PS)-

Synonyms: (R,R)-(2',3')c-diAM(PS)2,

3’3’-c(ApIp) sodium salt

2’FdIM(PS]) sodium salt

(2',3')-Rp,Rp-c-diAMPSS.

Formula: C20H23N9O13P2 .2Na

Formula: C20H21F2N9O9P2S2 .2Na

Formula: C20H22N10O10P2S2 .2Na.

Molecular weight: 659.4 (free acid)

Molecular weight: 695.5 (free acid)

Molecular weight: 734.50.

703.4 (sodium salt)

739.5 (sodium salt)

Purity: ≥ 95% by LC/MS & NMR.

Purity: ≥ 95% by LC/MS & NMR.

Purity: ≥ 95% by LC/MS & NMR.

Solubility: 50 mg/ml in water

Solubility: 50 mg/ml in water

Solubility: 50 mg/ml in water.

Cat Code

Cat Code

Cat Code

tlrl-nacai

tlrl-nacairs(500ug)(100ug)tlrl-nacda2r-01 (100ug)
tlrl-nacda2r

InvivoGen Patent : US20160362441; US15535864, WO/2017/186711(500ug)Aduro Biotech/Novartis

Phosphorothiotate modified molecules (CL656 and CL695) are more inflammatory thanphosphodiester-based compounds
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CL656 treatment leads to an adaptive immune response
CL656 cured mice

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CL656 synergizes with anti PD-1 regress tumor growth   
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CL656 treatment leads to a better B16 tumor regression
2.105 B16 Cells s.c. implantation
on C57Bl/6 mice

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STING and Cancer“Marker” in Tumor   

Product Box
Products
THP1-Dual™ cells
THP1-Dual™ KO-STING cells
ISD NakedCat.
thpd-nfis
thpd-kostg
tlrl-isdn

Assaf Marcus, Amy J. Mao, Monisha Lensink-Vasan, LeeAnn
Wang, Russell E. Vance, David H. Raulet. Tumor-Derived cGAMP
Triggers a STING-Mediated Interferon Response in Non-tumor
Cells to Activate the NK Cell Response. Immunity. 2018

STING and Cancer

ISRE

Check Point & STINGHER2 suppresses cytosolic DNA sensing

Product Box
Products
Poly(dA:dT)
2’3’-cGAMPHER2 protects cancer cells from STING-mediated antitumour immunityCat.
tlrl-patn-1
tlrl-nacga23-1

Shiying Wu, et al. HER2 recruits AKT1 to disrupt STING signalling and suppress
antiviral defence and antitumour immunity. Nature Cell Biology. 2019

STING and Cancer
CD8+ T cell independent
Lymphoma

Colorectal tumor

Colorectal tumor

Melanoma

MHC-I deficienttumor

Leukemia

Product Box
Product

Cat. code

2’3’-c-di-AM(PS)2 (Rp,Rp)

tlrl-nacda2r

2’3’-c-di-AM(PS)2 (Rp,Rp)
induced rejection of MHC-I
deficient tumors depends
on NK cells

Christopher J. Nicolai, et al. NK cells mediate clearance of CD8+ T cell–resistant tumors in response to STING agonists. Sci. Immunology. 2020

STING and Cancer
Microbiota
2015

2020

MC38 Tumor

MC38 Tumor

Oral Administration

BMDCs coculture
with OTI cellCD8+ T cells coculture
with MC38 cells

Bifidobacterium

A

Autoimmunity

cGAS
STING

Andrea Ablasser and Zhijian J. Chen. cGAS in action: Expanding roles in immunity and inflammation. Science. 2019

STING and Autoimmunity

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Product

Cat. code

THP1-Dual™ KO-TREX1 Cells

thpd-kotrex

THP1-Dual™ KO-cGAS Cells

thpd-kocgas

THP1-Dual™ KO-STING Cells

thpd-kostg

THP1-Dual™ KI-hSTING-S154 Cells

thpd-s154

THP1-Dual™ KI-hSTING-M155 Cells

thpd-m155

THP1-Dual™ KO-IRF3 Cells

thpd-koirf3
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STING and Autoimmunity

SAMHD1 and Aicardi-Goutieres Syndrome (AGS)SAMHD1 prevents accumulation of cytosolic ssDNA andinduction of type I IFNs mediated by cGAS-STING pathway.
Flavie Coquel, et. al. SAMHD1 acts at stalled replication forks to prevent interferon induction. Nature. 2018

STING and Autoimmunity

DNaseI1-/- and Polyarthritis

WT: DNase II/ Ifnar1/
DKO: DNase II/ STING/

Ahn, J., Gutman, D., Saijo, S. & Barber, G. N. STING manifests self DNA-dependent inflammatory disease.
Proc. Natl Acad. Sci. USA 109, 19386–19391 (2012).

STING and Autoimmunity

Trex1-/- andAicardi-Goutieres
Syndrome (AGS)
Product Box
Product
Elizabeth E. Gray, et al. Journal of Immunology. 2015

5'ppp-dsRNA

Cat. code
tlrl-3prna

ISD Naked tlrl-isdnSensorRIG-I
cGAS

THP1-Dual KO-TREX1 Cells thpd-kotrex

Daxing Gao, et al. Proc Natl Acad Sci. 2015

STING and Autoimmunity

Trex1-/-/ and AGS
Product Box
Cat. code
SensorThose data suggest new drugs block ProductcGAStlrl-3prna
at doseRIG-I
Elizabeth
Eli
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E. G
Gray, et al.l JJournall off IImmunology.
l2015
dependent manner might be ISD
efficaciouson
Naked tlrl-isdn
cGAS
THP1-Dual KO-TREX1
thpd-kotrex
autoimmune disease, and avoid the immunodeficiency
Cells
induced by impaired responses to viral DNA
5'ppp-dsRNA
5'5'ppp-dsRNAd RNADaxing Gao, et al. Proc Natl Acad Sci. 2015

STING and Autoimmunity

cGAS inhibitor

Inhibition of cGAS activity in RAW macrophage and BMDM cells froman Aicardi-Goutières Syndrome mouse model.

Product Box
Product
Identification of cGAS inhibitors

Jessica Vincent, et al. Small molecule inhibition of cGAS reduces interferon expression inprimary macrophages from autoimmune mice. Nature Communication.2017

ISD Naked
RAW-Lucia ISG
RAW-Lucia ISG-KO- TREX1
RAW-Lucia ISG-KO-cGAS
RU.521

Cat. code
tlrl-isdn
rawl-isg
rawl-kotrex
rawl-kocgas
inh-ru521

STING and Autoimmunity

cGAS inhibitor
Key features of G140:
G140 specifically inhibits human cGAS
in a dose-dependent manner.
G140 inhibits both cGAS-mediatedIRF and NF-B signaling.
Each lot of G140 is highly pure(>95%) and functionally tested.

Product
THP1-Dual cells
RAW-Lucia cells
Poly(I:C)
G140 [NEW]Cat. code
thpd-nfis
rawl-isg
tlrl-pic
inh-g140

Lodoe Lama, et al. Development of human cGASspecific small molecule inhibitors for repression of
dsDNA triggered interferon expression .Nature
Communication.2019

STING and Autoimmunity

cGAS inhibitor         Product
c-di-AMP

Cat. code
tlrl-nacdg

2'3'-cGAMP

tlrl-nacga23

Poly (I:C)

tlrl-pic

BX795

tlrl-bx7

Jiang Dai, et al. Acetylation Blocks cGAS Activity and Inhibits
Self-DNA-Induced Autoimmunity. Cell. 2019                     !"      !"STING and Autoimmunity
Stimulator of Interferon Genes (STING)–AssociatedVasculopathy with Onset in Infancy (SAVI).

Y.Liu, et al. Activated STING in a
Vascular and Pulmonary Syndtrome.
N ENGL J MED. 2014

STING Variants

AdvertisementKnock Out Reporter Cell Lines
STING
and Autoimmunity

M155

NEW !
STING SAVI Reporter Cells
InvivoGen provides a collection of STING variant
reporter cells derived from the human THP-1 monocytic
cell line. Among them, two express a STING gain-offunction (S154 and M155) responsible for the STINGassociated vasculopathy with onset in infancy (SAVI).
SAVI-patients display a single-point mutation in the

S154

STING protein leading to its constitutive activation andexcessive activation of ISGs. While S154 results from a denovo germline mutation1, M155 is an inherited mutation2.
STING SAVI reporter cells are convenient and powerfultools for antagonist screening in the STING pathway.

Products
THP1-Dual™ KI-hSTING-S154 Cells

Cat.
thpd-s154

THP1-Dual™ KI-hSTING-M155 Cells

thpd-m155

BX795

tlrl-bx7

STING and Autoimmunity

STING inhibitor

Product Box
Product

Cat. codec-di-AMP

tlrl-nacda

DMXAA

tlrl-dmx

5’ppp-dsRNA

tlrl-3prna-100

H-151

inh-h151

Simone M. Haag et al.Targeting STING with covalent small-molecule inhibitors. Nature. 2018

STING and Autoimmunity

STING inhibitor:The best STING inhibitor by far

ADU-S100

HAQ

ADU-S100

WT

Effect of H-151 on the IRF response of THP1-Dual™
cells to 2’3’-cGAMP.

3’3’-cGAMP FluorinatedSimone M. Haag et al.Targeting STING with covalent small-molecule inhibitors. Nature. 2018

SAVI

SAVI

STING and Autoimmunity

STING Variants

Hiroyasu Konno, et al. Pro-inflammation Associated with a Gain-ofFunction Mutation (R284S) in the Innate Immune Sensor STING.
Cell Reports. 2018

AMPK Inhibitors: Doxorubicin and GSK 690693

STING and Autoimmunity

STING Variant
STING Isoforms
Cat. CodehSTING
puno1-hstingwt
mSTING
puno1-mstingwt
hSTING-A162
puno1-hsting-a162
hSTING-A230
puno1-hsting-a230
hSTING-H232
puno1-hsting-h232
hSTING-HAQ
puno1-hsting-haq
hSTING-M155
puno1-hsting-m155
hSTING-N200
puno1-hsting-n200
mSTING-Gt
puno1-msting-gt

KO-STING cells transfected with WT or mutant STING and HEK-BlueTM ISG cells(parental) were stimulated with 10 mg/ml of 2’3’-cGAMP, 3’3’-cGAMP, c-di-GMP orDMXAA. After 24h incubation, the levels of IRF-induced SEAP were determinedusing QUANTI-BlueTM.

Contents
STING variants are provided in the pUNO1
plasmid as 20 μg of lyophilized DNA. Each
plasmid is supplied with 4 pouches of Fastmedia® Blas and 1 ml blasticidin at 10 mg/ml.

STING and Autoimmunity

NATE™

Nucleic Acid Transfection Enhancer NEW

InvivoGen now offers the NATE™ reagent, a nucleic acid transfection enhancer, to boost both transient and stabletransfection efficiencies in hard-to-transfect cell lines such as monocytes(THP-1) and macrophages(RAW 264.7).



Exogenous nucleic acids are detected bycytosolic sensors such as cGAS/STING,
AIM2 inflammasome, and LC3 mediatedautophagy.



The NATE™ reagent inhibits a number ofnucleic acid sensing pathways, thereby
protecting
exogenousDNAandfacilitating its expression.

STING and Autoimmunity

Increased DNA expression in NATE™ treated cells$!%$  )$  )! $" #$! Ć '

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NATE™ enhances transient DNA expression in human THP-1 monocytes:
Transfection of a ~3 kb GFP-expressing plasmid into THP-1 cells was performed using GeneXPlus without (a:top; b:yellow) or with
(a:bottom; b:green) the NATE™ reagent. After 48 hours, cells were visualized by fluorescence microscopy (a), and transfection efficiency wasmeasured using flow cytometry (b). Data are presented as a fold change normalized to the transfection efficiency without NATE™ reagent.

STING and Autoimmunity

NATE™

Increased rate of stably transfected cells with NATE™

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NATE™ increases the number of stable SEAP-expressing clones in murine RAW 264.7 macrophages:
Transfection of a ~10 kb SEAP-expressing plasmid into RAW 264.7 cells was performed using Lipofectamine® LTX, without (left) or with(right) the NATE™ reagent. After 10 days in selection with Blasticidin, the number of stables clones expressing SEAP (blue wells) was readily
visualized using QUANTI- Blue™ Solution detection reagent.

STING and Autoimmunity

Also
Available

Do you realize that antibiotics can stimulate
immune responses
Molecular patterns of microorganisms can stimulate immune
cells is common wisdom. Antibiotics are produced frommicroorganisms, therefore they may contain the component of
microorganisms. As an immune specialist, InvivoGen is the only
provider test microorganisms contamination in selective
antibiotics.

Selective Antibotics

Solvent

Blasticidin

HEPES

Zeocin™ [1,2]

HEPES

Puromycin

HEPES

G418

H2 O

Hygromycin B Gold™

HEPES

Phleomycin

HEPES

InvivoGen Trademark[1] Gatignol, A., Durand, H. & Tiraby, G. Bleomycin resistance conferred by a drug-binding protein. FEBS Lett. 230: 171–175. 1988.
[2] Mulsant P, Gatignol A, Dalens M, Tiraby G. Phleomycin resistance as a dominant selectable marker in CHO cells. Somatic Cell and Molecular
Genetics. 14 (3): 243–52. 1988.

STING and Autoimmunity

Phase

Agent

TargetCompany

DiscoverycCAS inhibitor

Inflammation,
autoimmunity,
neuroinflammation

IFM and Novartis

Discovery

STING inhibitor

Inflammation,
autoimmunity,
neuroinflammation

IFM and Novartis

Preclinical

SB11736 (Orally
available STING
antagonist)

Autoimmunity &Inflammatory
Diseases

Spring Bank

Discovery

STING inhibitor

Inflammation

CURADEV

Discovery

STING inhibitor

Autoimmune
disordersNimbus Therapeutics

A

cGAS
STING

Vaccine against intracellular pathogen
and cancer has been considered ascellular immunity. Adjuvant that initiate
antigen-specific effector and long-lived
memoryCD4 and CD8 T cell
responses is becoming a hot topic

Adjuvant

STING and Adjuvant
STING vs Influenza Vaccine Adjuvant
Intradermally (ID) / Intramuscularly (IM)

Product Box
Products

Cat.

AddaVax™
c-di-GMP VacciGrade™
2’3’-cGAMP VacciGrade™
3’3’-cGAMP VacciGrade™
MPLA-SM VacciGrade™

vac-adx-10
vac-cdg
vac-nacga23
vac-nacga
vac-mpla-cGAMP shows superior adjuvant effect on
cutaneous vaccination-cGAMP augments intradermally H5N1 vaccine
immunizationJi Wang, et al.    !. 2016

STING and Adjuvant
STING vs Influenza Vaccine Adjuvant

Susanne Herold and Leif-Erik Sander. Toward a universal flu vaccine. Science. 2020

Product Box
Products

Cat.

Poly (I:C)

tlrl-pic

Pam3CSK3

tlrl-pms

LumiKine™ Xpress mIFN-*4+2

luex-mifnbv2

Ji Wang, et al. Pulmonary surfactant–biomimetic nanoparticles potentiate
heterosubtypic influenza immunity. Science. 2020

STING and Adjuvant
STING & HSV

Morten K. Skouboe, Alice Knudsen, Line S.
Reinert, Cedric Boularan, Thierry Lioux,
Eric Perouzel, Martin K. Thomsen, Søren R.
Paludan. STING agonists enable antiviralcross-talk between human cells and confer
protection against genital herpes in mice.
PLOS Pathogens. 2018

2’3’-cGAM (PS)2 (Rp/Sp) (125 &g/mouse)
Cat Code: tlrl-nacga2srs

Systemic administration of STING agonists improves thesurvival of mice infected with HSV2

STING and Adjuvant

STING vs Anti-tumor Vaccine
OVA257-264)

OVA265-280

CDNs induced potent STING-dependent CD4+, CD8+, andT helper 1 (TH1)–biased humoral immunity that was specificfor the coformulated ovalbumin (OVA) antigens

Product Box
Product
2’3’-c-di-AM(PS)2 (Rp,Rp)

Cat. code
tlrl-nacda2r

ML RR-S2 CDA (ADU-S100) with PD-1 blockade
cures mice bearing established tumors

Fu J, et al. STING agonist formulated cancer vaccines can cure established tumors resistant to PD-1 blockade. Sci Transl Med. 2015.

STING and Adjuvant

VacciGradeTM Ligands as adjuvants for Th1 response

InvivoGen Adjuvant

VacciGradeTM
Preclinical grade
Guaranteed sterile
Tested for presence ofendotoxins

Th1

cGAMP Transfer
HIV particle

CX43 gap-junction

Cell-Cell
fusion

SLC19A1P2X7R channel

VRAC

cGAMP Transfer

A. Bridgeman, et al.Viruses transfer the antiviral second messenger cGAMP between cells. Science. 2015

cGAMP Transfer
Cell-Cell Fusion

Membrane Fusion-Enabled Spread of Innate Immunity
Requires Functional STING, but Not cGAS, in Target Cells

Shuting Xu , et al. cGAS-Mediated Innate Immunity
Spreads Intercellularly through HIV-1 Env-Induced
Membrane Fusion Sites.  *.2016

Mounting of Coculture-Induced Type I IFN Requires HIV-1
Env and CD4/Coreceptor-Mediated Membrane Fusion and
Direct Cell-Cell Contacts.

cGAMP Transfer

Gap-Junction - Connexin
Qing chen, et al. Carcinoma–astrocyte gap
junctions promote brain metastasis by
cGAMP transfer. Nature. 2016

-CX43-dependent phosphorylationof TBK1 and IRF3
-Cancer cells trigger astrocyte
cytokine release via CX43

-STING promote brain metastasis-Cancer cells transfer cGAMP toastrocytes via CX43.

cGAMP Transfer

2019

2019

SLC19A1 is folatetransporter

FDA approved drug
methotrexate andsulfasalazine block the
SLC19A1 channel

Product Box
Product
3'3'-cGAMP
2'3'-cGAMP
2’2'-cGAMP
2’3’-c-di-AM(PS)2 (Rp,Rp)

Cat. code
tlrl-nacga
tlrl-nacga23
tlrl-nacga22
tlrl-nacda2r

cGAMP Transfer

2020

Product Box
Product
5’ppp-dsRNA
2'3'-cGAMP
DMXAA
ATP

Cat. code
tlrl-3prna-100
tlrl-nacga23
tlrl-dmx
tlrl-atpl

The entry of tumor-cell-produced cGAMP into
host cells was facilitated by the ATP-gated P2X
purinoceptor 7 (P2X7) receptor (P2X7R).

cGAMP Transfer
2020

VRAC Channel Activity Positively Correlates with ExtracellularcGAMP Responses

Product Box
Product
3'3'-cGAMP
2'3'-cGAMPPolyI:C LMW
LPS-EK

Cat. code
tlrl-nacga
tlrl-nacga23-5
tlrl-picw
tlrl-eklps

LRRC8 VRACs Directly Transport cGAMPAndrea Ablasser and Zhijian J. Chen. cGAS in action:
Expanding roles in immunity and inflammation. Science. 2019

Comparative Potency of cGAS Ligands

G3-YSD is a 26-mer DNA sequence derived from the HIV-1 RNA genome. This
short DNA displays a Y-shape arising from its palindromic sequence flanked byunpaired guanosine trimers (G3). The guanosine overhangs in this Y-form DNA havebeen identified as minimal recognition motifs for cGAS (cyclic GMP-AMP synthase,cGAMP synthase), a critical cytosolic DNA sensor.
Herzner AM. et al., 2015. Sequence-specific activation of the DNA sensor cGAS by Y-form DNA structures as found inprimary HIV-1 cDNA. Nat Immunol. 16(10):1025-33.

VACV-70 is a 70 bp oligonucleotides containing poxviral genome DNA motifs.
VACV-70 derives from the vaccinia virus DNA. Transfected VACV-70 was shown topotently induce IFN-in a TLR-, DAI- and RNA Pol III-independent, but STING-,
TBK1- and IRF3-dependent manner.
Unterholzner L. et al., 2010. IFI16 is an innate immune sensor for intracellular DNA. Nat Immunol. 11(11):997-1004.

Product Name
G3-YSD
G3-YSD Control
VACV-70 Naked
VACV-70/LyoVecTM

Cat. code
tlrl-ydna
tlrl-ydnac
tlrl-vav70n
tlrl-vav70c

Endotoxin Free Ligands
Endotoxin: False positive
results
RIG-1, STING, TLRs, NOD1/NOD2,
Inflammasome
EndotoxinnBacterial cell

LPS

Mycoplasma
wwall compone
components

Flagellin

CD14

LBP
MD2

TLR4

TLR1

TIRAP
MyD88

dsRNA

TLR6

TLR2

TLR5

MyD88

TIRAP
MyD88

A

TLR4
TLR3

IRAK4
IRAK1 IRAK2
Pellino TRAF6 Bcl-10

TRAM
TRIF

MALT1

TRAF3
TANK NAP1

TRADD
RIP1

TAB2 TAB3
TAK1

TBK1 IKKε
DDX3

MKKs

NEMO
IKKα IKKβ

P

ERK

IRF3

P

P

JNK

p38

p50

IκB
p65

IRF3 IRF3

c-foss Jun

p50 p65

ISRE3

AP1

NF-κB

IFN-β

Pro-inflammatory cytokines

Biological Activity Tested
Product Box
Product
2'3'-cGAMP

Cat. code
tlrl-cga23

SensorSTING

TLR2,TLR4 Negative:
Avoid the contamination ofBacteria, Mycoplasma, Fungi

T
TLR4
Negative:
En
Endotoxin
Free

THP1-Blue™ Positive:
Biological activity confirmed

Anti-Contamination – Detection
RAW-Blue cells

HEK-Blue cells

FFDA
DA testedd TLR2/TLR4
TLR2/TLR4 contamination
contamiinatiion iinncommercial
i l IFNb productsdInvivoGen reporter cells suggest Betaseron contains
TLR2/4 impurities

Commercial LAL-based tests showed noendotoxin (TLR4) contamination

Mice experiments suggest InvivoGen cell lines
are accurate

InvivoGen cell lines detect low levels of
impurities that are not evident in LAL assay

Anti-Contamination

Since 1980s
Application

Product
Plasmocin™ prophylactic
Normocin™

Prevetion

Primocin™
Fungin™

Detection

PlasmoTest™
Plasmocin™ treatment
Plasmocure™

Elimination

Normocure™
Fungin™

Anti-Contamination – Prevention

Normocin™
Mycoplasma

Bacteria

Fungi

Normocin™



Pen/Strep 

Anti-Contamination – Elimination

Normocure™
Anti - Bacteria

HEK293 cells (3x105 cells/ml) were spiked with a mixture ofGram- non-fermenting bacilli (Pseudomonas aeruginosa,
Alcaligenes xylosodans, Achromobacter sp. and Stenotrophomonas
maltophilia) at the concentration of 105 colony forming units(cfu)/ml, and were then either left untreated, or treated with
100 U/ml penicillin and 100 µg/ml streptomycin or with 100µg/ml Normocure™, After 4 days at 37C, 5% CO2, thebacteria were quantified (cfu per ml).

SARS-CoV-2 Related Products

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tech.hk@invivogen.com