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| Status |
Public on Aug 22, 2022 |
| Title |
Bioorthogonal Chemical Epigenetic Modifiers Enable Dose-dependent CRISPR Targeted Gene Activation |
| Organism |
Homo sapiens |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
CRISPR-Cas9 systems have been developed to regulate gene expression by using either fusions to epigenetic regulators or, more recently, through the use of chemically mediated strategies. These approaches have armed researchers with new tools to examine the function of proteins by intricately controlling expression levels of specific genes. Here, we present a CRISPR based chemical approach that uses a new Chemical Epigenetic Modifier (CEM) to hone to a gene targeted with a catalytically inactive Cas9 (dCas9) bridged to an FK506-binding protein (FKBP). One arm of the bifunctional CEM recruits BRD4 to the target site and the other arm is composed of a bumped ligand that binds to a mutant FKBP with a compensatory hole at F36V. This bump-and-hole strategy allows for activation of target genes in a dose dependent and reversible fashion with increased specificity and high efficacy.
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| |
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| Overall design |
ChIP-seq for BRD4 in conditions sgNT, sgCXCR4, 25nM CEM207, and sgCXCR4+25nM CEM207 to analyze the specificity of BRD4 binding in each.
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| |
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| Contributor(s) |
Lu D, Foley CA, Birla SV, Hepperla AJ, Simon JM, James LI, Hathaway NA |
| Citation(s) |
35302756 |
| |
| Submission date |
Oct 14, 2021 |
| Last update date |
Aug 25, 2022 |
| Contact name |
Austin J Hepperla |
| E-mail(s) |
hepperla@unc.edu
|
| Organization name |
University of North Carolina at Chapel Hill
|
| Department |
Genetics
|
| Street address |
7018B Mary Ellen Jones Building
|
| City |
Chapel Hill |
| State/province |
NC |
| ZIP/Postal code |
27599 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
|
| Samples (16)
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|
| Relations |
| BioProject |
PRJNA771351 |
| SRA |
SRP341405 |
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