Genomic targeting of epigenetic probes using a chemically tailored Cas9 system.

TitleGenomic targeting of epigenetic probes using a chemically tailored Cas9 system.
Publication TypeJournal Article
Year of Publication2017
AuthorsLiszczak, GP, Brown, ZZ, Kim, SH, Oslund, RC, David, Y, Muir, TW
JournalProc Natl Acad Sci U S A
Volume114
Issue4
Pagination681-686
Date Published2017 Jan 24
ISSN1091-6490
Abstract

Recent advances in the field of programmable DNA-binding proteins have led to the development of facile methods for genomic localization of genetically encodable entities. Despite the extensive utility of these tools, locus-specific delivery of synthetic molecules remains limited by a lack of adequate technologies. Here we combine the flexibility of chemical synthesis with the specificity of a programmable DNA-binding protein by using protein trans-splicing to ligate synthetic elements to a nuclease-deficient Cas9 (dCas9) in vitro and subsequently deliver the dCas9 cargo to live cells. The versatility of this technology is demonstrated by delivering dCas9 fusions that include either the small-molecule bromodomain and extra-terminal family bromodomain inhibitor JQ1 or a peptide-based PRC1 chromodomain ligand, which are capable of recruiting endogenous copies of their cognate binding partners to targeted genomic binding sites. We expect that this technology will allow for the genomic localization of a wide array of small molecules and modified proteinaceous materials.

DOI10.1073/pnas.1615723114
Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID28069948
PubMed Central IDPMC5278450