TY - JOUR
T1 - Reprogramming cell fate with a genome-scale library of artificial transcription factors
AU - Eguchi, Asuka
AU - Wleklinski, Matthew J.
AU - Spurgat, Mackenzie C.
AU - Heiderscheit, Evan A.
AU - Kropornicka, Anna S.
AU - Vu, Catherine K.
AU - Bhimsaria, Devesh
AU - Swanson, Scott A.
AU - Stewart, Ron
AU - Ramanathan, Parameswaran
AU - Kamp, Timothy J.
AU - Slukvin, Igor
AU - Thomson, James A.
AU - Dutton, James R.
AU - Ansari, Aseem Z.
N1 - Publisher Copyright:
© 2016, National Academy of Sciences. All rights reserved.
PY - 2016/12/20
Y1 - 2016/12/20
N2 - Artificial transcription factors (ATFs) are precision-tailored molecules designed to bind DNA and regulate transcription in a preprogrammed manner. Libraries of ATFs enable the high-throughput screening of gene networks that trigger cell fate decisions or phenotypic changes. We developed a genome-scale library of ATFs that display an engineered interaction domain (ID) to enable cooperative assembly and synergistic gene expression at targeted sites. We used this ATF library to screen for key regulators of the pluripotency network and discovered three combinations of ATFs capable of inducing pluripotency without exogenous expression of Oct4 (POU domain, class 5, TF 1). Cognate site identification, global transcriptional profiling, and identification of ATF binding sites reveal that the ATFs do not directly target Oct4; instead, they target distinct nodes that converge to stimulate the endogenous pluripotency network. This forward genetic approach enables cell type conversions without a priori knowledge of potential key regulators and reveals unanticipated gene network dynamics that drive cell fate choices.
AB - Artificial transcription factors (ATFs) are precision-tailored molecules designed to bind DNA and regulate transcription in a preprogrammed manner. Libraries of ATFs enable the high-throughput screening of gene networks that trigger cell fate decisions or phenotypic changes. We developed a genome-scale library of ATFs that display an engineered interaction domain (ID) to enable cooperative assembly and synergistic gene expression at targeted sites. We used this ATF library to screen for key regulators of the pluripotency network and discovered three combinations of ATFs capable of inducing pluripotency without exogenous expression of Oct4 (POU domain, class 5, TF 1). Cognate site identification, global transcriptional profiling, and identification of ATF binding sites reveal that the ATFs do not directly target Oct4; instead, they target distinct nodes that converge to stimulate the endogenous pluripotency network. This forward genetic approach enables cell type conversions without a priori knowledge of potential key regulators and reveals unanticipated gene network dynamics that drive cell fate choices.
KW - Artificial transcription factor
KW - Cell fate
KW - Gene regulatory networks
KW - Genome-scale library
KW - Reprogramming
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U2 - 10.1073/pnas.1611142114
DO - 10.1073/pnas.1611142114
M3 - Article
C2 - 27930301
AN - SCOPUS:85006340254
SN - 0027-8424
VL - 113
SP - E8257-E8266
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 51
ER -