@article{b2e6e83229184ed89520dc245727ed10,
title = "A protocol for rapid degradation of endogenous transcription factors in mammalian cells and identification of direct regulatory targets",
abstract = "Transcriptional changes happen within minutes; however, RNAi or genetic deletion requires days to weeks before transcription networks can be analyzed. This limitation has made it challenging to distinguish direct from indirect targets of sequence-specific transcription factors. This inability to define direct transcriptional targets hinders detailed studies of transcriptional mechanisms. This protocol combines rapid degradation of endogenous transcription factors with nascent transcript analysis to define the earliest, and likely direct, regulatory targets of transcription factors. For complete details on the use and execution of this protocol, please refer to Stengel et al., 2021).",
keywords = "CRISPR, Cell Biology, Flow Cytometry/Mass Cytometry, Genomics, Molecular Biology, Protein Biochemistry",
author = "Layden, {Hillary M.} and Eleuteri, {Nicholas A.} and Hiebert, {Scott W.} and Stengel, {Kristy R.}",
note = "Funding Information: We thank the members of the Hiebert lab for helpful discussions, reagents, and advice. We thank the VICC Flow Cytometry, Chemical Synthesis, and Genome Sciences Shared Resources for services and support. This work was supported by the T.J. Martell Foundation , the Robert J. Kleberg, Jr. and Helen C. Kleberg Foundation , the National Institutes of Health (grants RO1- CA178030 , RO1-CA164605 , and R01-CA64140 ), core services performed through Vanderbilt Digestive Disease Research grant NIDDK P30DK58404 and Vanderbilt-Ingram Cancer Center support grant NCI P30CA68485 , and the National Center for Advancing Translational Sciences (grant 2 UL1 TR000445-06 ). H.M.L. and K.R.S. were supported by NCI grant 5 T32 CA009582 . H.M.L. was supported by NHBLI NRSA fellowship F31HL156565 . K.R.S. was supported by American Cancer Society postdoctoral fellowship PF-13-303-01-DMC . Publisher Copyright: {\textcopyright} 2021 The Author(s)",
year = "2021",
month = jun,
day = "18",
doi = "10.1016/j.xpro.2021.100530",
language = "English (US)",
volume = "2",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "2",
}