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Related Experiment Video

Updated: Aug 15, 2025

Site Specific Lysine Acetylation of Histones for Nucleosome Reconstitution using Genetic Code Expansion in Escherichia coli

Site Specific Lysine Acetylation of Histones for Nucleosome Reconstitution using Genetic Code Expansion in Escherichia coli

Published on: December 26, 2020

Computational design of constitutively active cGAS.

Quinton M Dowling^1,2, Hannah E Volkman³, Elizabeth E Gray^3,4

¹Department of Bioengineering, University of Washington, Seattle, WA, USA.

Nature Structural & Molecular Biology

|January 2, 2023

Summary

Scientists engineered a constitutively active form of cyclic GMP-AMP synthase (cGAS) that triggers an immune response without DNA. This DNA-independent cGAS (CA-cGAS) showed therapeutic potential by reducing tumors in vivo.

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Area of Science:

Biochemistry
Immunology
Structural Biology

Background:

Cyclic GMP-AMP synthase (cGAS) is a key innate immune sensor activated by double-stranded DNA (dsDNA).
Activated cGAS produces cGAMP, stimulating the STING pathway and interferon production, crucial for defense against pathogens and cancer.

Purpose of the Study:

To design constitutively active cGAS (CA-cGAS) that is independent of dsDNA binding.
To validate the structural and functional properties of CA-cGAS.
To assess the therapeutic potential of CA-cGAS in vivo.

Main Methods:

Utilized two-state computational design and informatics-guided design to engineer CA-cGAS variants.
Confirmed DNA-independent active conformation using X-ray crystallography.
Evaluated in vivo efficacy by expressing CA-cGAS in tumor cells.

Main Results:

Developed CA-cGAS mutants exhibiting significant interferon-stimulating activity, comparable to dsDNA-stimulated wild-type cGAS.
X-ray crystallography confirmed the constitutive active conformation of CA-cGAS.
In vivo studies demonstrated STING-dependent tumor regression upon CA-cGAS expression in tumor cells.

Conclusions:

Successfully designed DNA-independent active cGAS (CA-cGAS) through computational and informatics-guided approaches.
CA-cGAS exhibits therapeutic potential, inducing STING-dependent tumor regression.
CA-cGAS variants offer a framework for enzyme stabilization and can serve as adjuvants or research tools for inflammatory diseases.