Title | Genomic targeting of epigenetic probes using a chemically tailored Cas9 system. |
Publication Type | Journal Article |
Year of Publication | 2017 |
Authors | Liszczak, GP, Brown, ZZ, Kim, SH, Oslund, RC, David, Y, Muir, TW |
Journal | Proc Natl Acad Sci U S A |
Volume | 114 |
Issue | 4 |
Pagination | 681-686 |
Date Published | 2017 Jan 24 |
ISSN | 1091-6490 |
Keywords | Amino Acid Sequence, Binding Sites, CRISPR-Associated Proteins, CRISPR-Cas Systems, DNA-Binding Proteins, Epigenomics, Escherichia coli, Genomics, Recombinant Proteins |
Abstract | <p>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.</p> |
DOI | 10.1073/pnas.1615723114 |
Alternate Journal | Proc Natl Acad Sci U S A |
PubMed ID | 28069948 |
PubMed Central ID | PMC5278450 |
Grant List | F32 GM110880 / GM / NIGMS NIH HHS / United States P01 CA196539 / CA / NCI NIH HHS / United States R01 GM107047 / GM / NIGMS NIH HHS / United States R37 GM086868 / GM / NIGMS NIH HHS / United States |