RT Journal Article
SR Electronic
T1 A viral toolkit for recording transcription factor-DNA interactions in live mouse tissues
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 538504
DO 10.1101/538504
A1 Alexander J. Cammack
A1 Arnav Moudgil
A1 Tomas Lagunas
A1 Jiayang Chen
A1 Michael J. Vasek
A1 Mark Shabsovich
A1 Katherine McCullough
A1 June He
A1 Xuhua Chen
A1 Misha Hooda
A1 Michael N. Wilkinson
A1 Timothy M. Miller
A1 Robi D. Mitra
A1 Joseph D. Dougherty
YR 2019
UL http://biorxiv.org/content/early/2019/10/17/538504.abstract
AB 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 endpoint 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, termed Flip-Excision (FLEX) calling cards 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.