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. 2023 Jul 10;120(29):e2214320120. doi: 10.1073/pnas.2214320120

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Copyright © 2023 the Author(s). Published by PNAS.

This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 1. — Comb-structured mRNA with immunostimulatory dsRNA teeth. This formulation is prepared from in vitro transcribed RNA with 5′ triphosphate (5′ppp-RNA), mRNA, and a chemically synthesized counter-RNA strand complementary to the 5′ppp-RNA and particular sequences in mRNA (cRNA). dsRNA teeth from 5′ppp-RNA and cRNA possess blunt-ended 5′ triphosphate, a substrate of RIG-I. The length of the cRNA sequence complementary to mRNA is fixed to 17 nt to maintain mRNA translational activity. In addition, design parameters of comb-structured mRNA, including the lengths and sequences of dsRNA teeth, and lengths of gap sequence in cRNA between two regions complementary to 5′ppp-RNA and mRNA, were optimized.