A modular dCas9-based recruitment platform for combinatorial epigenome editing

Tessa Swain, Christian Pflueger, Saskia Freytag, Daniel Poppe, Jahnvi Pflueger, Trung Viet Nguyen, Ji Kevin Li, Ryan Lister

Research output: Contribution to journalArticlepeer-review

Abstract

Targeted epigenome editing tools allow precise manipulation and investigation of genome modifications, however they often display high context dependency and variable efficacy between target genes and cell types. While systems that simultaneously recruit multiple distinct 'effector' chromatin regulators can improve efficacy, they generally lack control over effector composition and spatial organisation. To overcome this we have created a modular combinatorial epigenome editing platform, called SSSavi. This system is an interchangeable and reconfigurable docking platform fused to dCas9 that enables simultaneous recruitment of up to four different effectors, allowing precise control of effector composition and spatial ordering. We demonstrate the activity and specificity of the SSSavi system and, by testing it against existing multi-effector targeting systems, demonstrate its comparable efficacy. Furthermore, we demonstrate the importance of the spatial ordering of the recruited effectors for effective transcriptional regulation. Together, the SSSavi system enables exploration of combinatorial effector co-recruitment to enhance manipulation of chromatin contexts previously resistant to targeted editing.

Original languageEnglish
Article numbergkad1108
Pages (from-to)474-491
Number of pages18
JournalNucleic Acids Research
Volume52
Issue number1
Early online date24 Nov 2023
DOIs
Publication statusPublished - 11 Jan 2024

Cite this