Table of Contents

1. Intercellular communication in the innate immune system through the cGAS-STING pathway
Ramya Nandakumar, Samuel Joseph Windross and Søren R. Paludan
2. Analysis of human cGAS activity and structure
Wen Zhou, Aaron T. Whiteley and Philip J. Kranzusch
3. Chemical synthesis, purification, and characterization of 3′-5′-linked canonical cyclic dinucleotides (CDNs)
Changhao Wang, Min Hao, Qianqian Qi, Yashao Chen and Jörg S. Hartig
4. An SPR-based analysis of cGAS substrate KD and steady-state KM values
Erik C. Ralph and Justin Hall
5. A pyrophosphatase-coupled assay to monitor the NTase activity of cGAS
Richard Hooy and Jungsan Sohn
6. Tracking the polymerization of DNA sensors and inflammasomes using FRET
Zachary Mazanek and Jungsan Sohn
7. Monitoring gasdermin pore formation in vitro
Shiyu Xia, Jianbin Ruan and Hao Wu
8. Measuring TREX1 and TREX2 exonuclease activities
Wayne O. Hemphill and Fred W. Perrino
9. Kinetics of DNA: Protein association and dissociation by stopped-flow spectroscopy
Johannes Rudolph and Karolin Luger
10. Characterization of DNA bound cyclic GMP-AMP synthase using atomic force microscopy imaging
Alexander Lushnikov, Richard Hooy, Jungsan Sohn and Alexey Krasnoslobodtsev
11. Preparation of filamentous proteins for electron microscopy visualization and reconstruction
Mariusz Matyszewski and Jungsan Sohn
12. Cryo-EM studies of NAIP–NLRC4 inflammasomes
Nicole Haloupek, Patricia Grob, Jeannette Tenthorey, Russell E. Vance and Eva Nogales
13. An integrative protocol for the structure determination of the mouse ASC-PYD filament
Adam Mazur, Petr Broz and Sebastian Hiller
14. Protein interactions of the inflammasome adapter ASC by solution NMR
Pedro Diaz-Parga and Eva de Alba
15. Tracking DNA sensor and inflammasome complexes in cells using confocal microscopy
Brendan Antiochos
16. Assessment of inflammasome and type I IFN responses to DNA viruses and DNA PAMPS
Guido Massaccesi and Michael A. Chattergoon
17. Biochemical and microscopic analysis of inflammasome complex formation
Patrycja Swacha, Nelson Ong’ondo Gekara and Saskia F. Erttmann
18. Comet and micronucleus assays for analyzing DNA damage and genome integrity
Hui Jiang and Nelson Ong'ondo Gekara
19. Workflows and considerations for investigating protein interactions of viral DNA sensors
Timothy R. Howard, Bokai Song and Ileana M. Cristea
20. TUBE and UbiCRest assays for elucidating polyubiquitin modifications in protein complexes
Swarupa Panda and Nelson Ong'ondo Gekara

About the Author

Jungsan Sohn is at Johns Hopkins School of Medicine, MD, USA