Programming DNA Circuits for Controlled Immunostimulation through CpG Oligodeoxynucleotide Delivery

Journal of the American Chemical Society, vol. 145, pp. 12465–12474

Abstract

Herein, we present a DNA circuit programmed for the delivery of CpG oligodeoxynucleotides (CpG ODNs) with the pharmacological immunostimulation function. The circuit employs a complementary DNA (cDNA) strand to deactivate the biological function of CpG ODNs via hybridization, while T7 exonuclease mediates the activation by hydrolyzing the cDNA and releasing the CpG ODN as an active moiety. We investigated the influence of several factors on the kinetic profile and temporal behavior of the circuit. These include the design of the cDNA strand, the concentration of the DNA duplex, and the concentration of T7 exonuclease. The DNA circuit's in vitro activation resulted in toll-like receptor 9 stimulation in the HEK-engineered cell line, as well as tumor necrosis factor-alpha release by J774A.1 macrophages. By programming the DNA circuit to control the release of the CpG ODN, we achieved an altered pharmacological profile with acute and potent immunostimulation, in comparison to a system without controlled CpG ODN release, which exhibited a slow and delayed response. Our findings demonstrate the potential of DNA circuits in controlling the pharmacological activity of DNA strands for controlled drug delivery.

Authors 3

1. Aman Ishaqat Aachen DWI - Leibniz Institute for Interactive Materials Institute of Technical and Macromolecular Chemistry

   RWTH Aachen University · DWI – Leibniz Institute for Interactive Materials

   Affiliation as printed

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstr. 50, 52074 Aachen, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 2, 52074 Aachen, Germany

2. Qing Liu

   Wenzhou Institute, University of Chinese Academy of Sciences

   Affiliation as printed

   Wenzhou Institute, University of Chinese Academy of Sciences, 25001 Wenzhou, China

3. Lifei Zheng corresponding

   Wenzhou Institute, University of Chinese Academy of Sciences

   Affiliation as printed

   Wenzhou Institute, University of Chinese Academy of Sciences, 25001 Wenzhou, China