Genomic targeting of epigenetic probes using a chemically tailored Cas9 system

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Abstract
Significance: Programmable DNA-binding proteins such as nuclease-deficient Cas9 (dCas9) offer a simple system for genomic localization of genetically encodable biomolecules. These tools have enabled characterization of numerous different chromatin effectors at specific genetic elements. The delivery of fully synthetic cargos to specific loci is currently limited by a lack of adequate technologies. Here we used protein trans -splicing to ligate chemical moieties to dCas9 in vitro and subsequently deliver these molecules to live cells. We demonstrate the effectiveness and versatility of this method by delivering both small-molecule and proteinaceous epigenetic probes to endogenous genomic sites where they interact with their target proteins. This facile and modular strategy will find broad use in locus-specific targeting of chemical cargos to study nuclear processes.
Funding Information
  • HHS | National Institutes of Health (F32GM110880)
  • HHS | National Institutes of Health (R37-GM086868)
  • HHS | National Institutes of Health (R01-GM107047)

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