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Author Notes:

Philip J. Santangelo, Philip.j.santangeloemory.edu

J.P.B., S.B., and P.J.S. conceived the study and designed the experiments. J.P.B., S.B., H.E.P., R.A.H., and J.A.W. performed the experiments. H.E.P. produced the mRNA. J.P.B. and J.A.W. formulated LNPs. N.I. and C.A.G. conceived of, created, and validated the VPH-dCas9-SS18 construct. J.P.B., S.B., and P.J.S. wrote the manuscript. J.P.B., S.B., N.I., C.Z., J.A.W., P.J.S., and C.A.G. edited the manuscript. J.P.B., S.B., and N.I. contributed equally to this work.

The authors declare the following competing financial interest(s): All authors declare no competing interests related to this work. NI and CAG are inventors on patent applications regarding CRISPR technologies and targeted gene activation. CAG is an advisor to Tune Therapeutics, Sarepta Therapeutics, and Levo Therapeutics, and a co-founder of Tune Therapeutics and Locus Biosciences.

Subject:

Research Funding:

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:

  • Science & Technology
  • Physical Sciences
  • Technology
  • Chemistry, Multidisciplinary
  • Chemistry, Physical
  • Nanoscience & Nanotechnology
  • Materials Science, Multidisciplinary
  • Chemistry
  • Science & Technology - Other Topics
  • Materials Science
  • CRISPR
  • dCas9
  • gene activation
  • mRNA
  • lipid nanoparticle
  • nonviral
  • in vivo
  • LIPID NANOPARTICLES
  • ENDOGENOUS GENES
  • EXPRESSION
  • MICE

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

Tools:

Journal Title:

ACS NANO

Volume:

Volume 16, Number 4

Publisher:

, Pages 5660-5671

Type of Work:

Article | Final Publisher PDF

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.

Copyright information:

© 2022 The Authors. Published by American Chemical Society

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/rdf).
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