Reprogramming cell fate with a genome-scale library of artificial transcription factors
| Autor: | Matthew J. Wleklinski, James A. Thomson, Evan A. Heiderscheit, Aseem Z. Ansari, James R. Dutton, Anna S. Kropornicka, Devesh Bhimsaria, Catherine K. Vu, Mackenzie C. Spurgat, Scott Swanson, Timothy J. Kamp, Ron Stewart, Asuka Eguchi, Igor I. Slukvin, Parameswaran Ramanathan |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Transcription Genetic viruses genetic processes Gene regulatory network Artificial transcription factor Computational biology Biology Cell fate determination Protein Engineering environment and public health Epigenesis Genetic Mice 03 medical and health sciences Protein Domains Transcription (biology) Animals Humans Cell Lineage Gene Regulatory Networks Transcription factor Zinc finger Genetics Genomic Library Binding Sites Multidisciplinary POU domain Sequence Analysis RNA fungi Zinc Fingers Fibroblasts Cellular Reprogramming Gene Expression Regulation Neoplastic HEK293 Cells 030104 developmental biology PNAS Plus Reprogramming Chaperonin Containing TCP-1 Transcription Factors |
| Zdroj: | Proceedings of the National Academy of Sciences. 113 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1611142114 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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