Harnessing CRISPR-Cas9 for Epigenetic Engineering

Rosa S Guerra-Resendez; Isaac B Hilton

doi:10.1007/978-1-0716-2421-0_14

Harnessing CRISPR-Cas9 for Epigenetic Engineering

Methods Mol Biol. 2022:2518:237-251. doi: 10.1007/978-1-0716-2421-0_14.

Authors

Rosa S Guerra-Resendez¹, Isaac B Hilton^{2

3

4}

Affiliations

¹ Systems, Synthetic, and Physical Biology PhD Program, Rice University, Houston, TX, USA.
² Systems, Synthetic, and Physical Biology PhD Program, Rice University, Houston, TX, USA. [email protected].
³ Department of Bioengineering, Rice University, Houston, TX, USA. [email protected].
⁴ Department of BioSciences, Rice University, Houston, TX, USA. [email protected].

PMID: 35666449
DOI: 10.1007/978-1-0716-2421-0_14

Abstract

Epigenome editing has become more precise and effective by coupling epigenetic effectors to the dCas9 protein and targeting regulatory regions such as promoters and enhancers. Here, we describe a basic methodology for performing an epigenome editing experiment, starting from gRNA design and cloning to transiently transfecting the gRNA plasmid and the CRISPR/dCas9-based epigenetic effector and finalizing with chromatin immunoprecipitation (ChIP) to validate changes in epigenetic state at a targeted genomic region.

Keywords: CRISPR; Deactivated Cas9; Epigenome editing; Guide RNA; Synthetic biology.

MeSH terms

CRISPR-Cas Systems* / genetics
Epigenome
Epigenomics
Gene Editing / methods
RNA, Guide, CRISPR-Cas Systems* / genetics
RNA, Guide, CRISPR-Cas Systems* / metabolism

Substances

RNA, Guide, CRISPR-Cas Systems