Research Article | Open Access

Volume 2021 |Article ID 9815820 | https://doi.org/10.34133/2021/9815820

Durable CRISPR-Based Epigenetic Silencing

Muneaki Nakamura,¹ Alexis E. Ivec,^1,2 Yuchen GaoiD ,^1,3 Lei S. Qi iD ^1,4,5

¹Department of Bioengineering, Stanford, CA 94305, USA
²Program in Human Biology, Stanford, CA 94305, USA
³Cancer Biology Program, Stanford, CA 94305, USA
⁴Department of Chemical and Systems Biology, Stanford, CA 94305, USA
⁵ChEM-H Institute, Stanford, CA 94305, USA

Abstract

Development of CRISPR-based epigenome editing tools is important for the study and engineering of biological behavior. Here, we describe the design of a reporter system for quantifying the ability of CRISPR epigenome editors to produce a stable gene repression. We characterize the dynamics of durable gene silencing and reactivation, as well as the induced epigenetic changes of this system. We report the creation of single-protein CRISPR constructs bearing combinations of three epigenetic editing domains, termed KAL, that can stably repress the gene expression. This system should allow for the development of novel epigenome editing tools which will be useful in a wide array of biological research and engineering applications.