A viral toolkit for recording transcription factor–DNA interactions in live mouse tissues
| Autor: | Robi D. Mitra, Katherine B. McCullough, Susan E. Maloney, June He, Allen Yen, Mark Shabsovich, Joseph D. Dougherty, Michael J. Vasek, Arnav Moudgil, Jiayang Chen, Tomas Lagunas, Timothy M. Miller, Alexander J. Cammack, Michael N. Wilkinson, Misha Hooda, Xuhua Chen |
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| Rok vydání: | 2020 |
| Předmět: |
Epigenomics
Cell type Cell Computational biology Biology Genome Antibodies Mice medicine Animals Humans Tissue Distribution Epigenetics Enhancer Transcription factor Regulation of gene expression Binding Sites Multidisciplinary Integrases Biological Sciences Dependovirus Chromatin DNA-Binding Proteins medicine.anatomical_structure Gene Expression Regulation DNA Transposable Elements Chromatin immunoprecipitation Algorithms Transcription Factors |
| Zdroj: | Proc Natl Acad Sci U S A |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1918241117 |
| Popis: | Transcription factors (TFs) enact precise regulation of gene expression through site-specific, genome-wide binding. Common methods for TF-occupancy profiling, such as chromatin immunoprecipitation, are limited by requirement of TF-specific antibodies and provide only end-point snapshots of TF binding. Alternatively, TF-tagging techniques, in which a TF is fused to a DNA-modifying enzyme that marks TF-binding events across the genome as they occur, do not require TF-specific antibodies and offer the potential for unique applications, such as recording of TF occupancy over time and cell type specificity through conditional expression of the TF–enzyme fusion. Here, we create a viral toolkit for one such method, calling cards, and demonstrate that these reagents can be delivered to the live mouse brain and used to report TF occupancy. Further, we establish a Cre-dependent calling cards system and, in proof-of-principle experiments, show utility in defining cell type-specific TF profiles and recording and integrating TF-binding events across time. This versatile approach will enable unique studies of TF-mediated gene regulation in live animal models. |
| Databáze: | OpenAIRE |
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