Tailoring a CRISPR/Cas-based Epigenome Editor for Programmable Chromatin Acylation and Decreased Cytotoxicity.

Autor:	Goell J; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Li J; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Mahata B; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Ma AJ; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Kim S; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Shah S; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Shah S; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Contreras M; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Misra S; Department of Biosciences, Rice University, Houston, TX 77030, USA., Reed D; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Bedford GC; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Escobar M; Department of Bioengineering, Rice University, Houston, TX 77030, USA., Hilton IB; Department of Bioengineering, Rice University, Houston, TX 77030, USA.; Department of Biosciences, Rice University, Houston, TX 77030, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Sep 22. Date of Electronic Publication: 2024 Sep 22.
DOI:	10.1101/2024.09.22.611000
Abstrakt:	Engineering histone acylation states can inform mechanistic epigenetics and catalyze therapeutic epigenome editing opportunities. Here, we developed engineered lysine acyltransferases that enable the programmable deposition of acetylation and longer-chain acylations. We show that targeting an engineered lysine crotonyltransferase results in weak levels of endogenous enhancer activation yet retains potency when targeted to promoters. We further identify a single mutation within the catalytic core of human p300 that preserves enzymatic activity while substantially reducing cytotoxicity, enabling improved viral delivery. We leveraged these capabilities to perform single-cell CRISPR activation screening and map enhancers to the genes they regulate in situ . We also discover acylation-specific interactions and find that recruitment of p300, regardless of catalytic activity, to prime editing sites can improve editing efficiency. These new programmable epigenome editing tools and insights expand our ability to understand the mechanistic role of lysine acylation in epigenetic and cellular processes and perform functional genomic screens. Competing Interests: DECLARATION OF INTERESTS J.G. and I.B.H are inventors on patents related to this work. J.G., J.L., B.M., M.E., and I.B.H. are inventors on patents related to genome and epigenome editing technologies. J.G. and I.B.H. are founders of Mercator Biosciences.
Databáze:	MEDLINE
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