Robust, Durable Gene Activation In Vivo via mRNA-Encoded Activators

• Type of Material: Article
• Authors: Jared P Beyersdorf, Georgia Institute of Technology; Swapnil Bawage, Georgia Institute of Technology; Nahid Iglesias, Duke University; Hannah E Peck, Georgia Institute of Technology; Ryan A Hobbs, Georgia Institute of Technology; Jay A Wroe, Georgia Institute of Technology; Chiara Zurla, Georgia Institute of Technology; Charles A Gersbach, Duke University; Philip Santangelo, Emory University
• Language: English
• Date: 2022-03-31
• Publisher: AMER CHEMICAL SOC
• Publication Version: Final Published Version
• Copyright Statement: © 2022 The Authors. Published by American Chemical Society
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1021/acsnano.1c10631
• Title of Journal or Parent Work: ACS NANO
• Volume: 16
• Issue: 4
• Start Page: 5660
• End Page: 5671
• Grant/Funding Information: This study was supported by the Defense Advanced Research Projects Agency, Grant No. HR00111920008, National Institutes of Health Grant No. U01AI146356, National Science Foundation Grant No. EFMA-1830957, and an Allen Distinguished Investigator Award to C.A.G.
• Keywords: Chemistry, Multidisciplinary; Materials Science, Multidisciplinary; Materials Science; mRNA; Chemistry, Physical; Nanoscience & Nanotechnology; dCas9; Chemistry; Science & Technology - Other Topics; lipid nanoparticle; Science & Technology; gene activation; ENDOGENOUS GENES; CRISPR; EXPRESSION; Physical Sciences; Technology; MICE; nonviral; LIPID NANOPARTICLES; in vivo
• Research Categories: Engineering, Biomedical

Abstract

Programmable control of gene expression via nuclease-null Cas9 fusion proteins has enabled the engineering of cellular behaviors. Here, both transcriptional and epigenetic gene activation via synthetic mRNA and lipid nanoparticle delivery was demonstrated in vivo. These highly efficient delivery strategies resulted in high levels of activation in multiple tissues. Finally, we demonstrate durable gene activation in vivo via transient delivery of a single dose of a gene activator that combines VP64, p65, and HSF1 with a SWI/SNF chromatin remodeling complex component SS18, representing an important step toward gene-activation-based therapeutics. This induced sustained gene activation could be inhibited via mRNA-encoded AcrIIA4, further improving the safety profile of this approach.