Cell Stem Cell
Volume 25, Issue 4, 3 October 2019, Pages 486-500.e9
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Article
Mitigating Antagonism between Transcription and Proliferation Allows Near-Deterministic Cellular Reprogramming

https://doi.org/10.1016/j.stem.2019.08.005Get rights and content
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Highlights

  • Chemical, genetic cocktail reduces genomic stress induced by TF reprogramming

  • DDRR cocktail expands population of hypertranscribing, hyperproliferating cells (HHCs)

  • Supported by topoisomerases, HHCs reprogram at near-deterministic rates

  • Topoisomerase expression reduces negative DNA supercoiling and R-loop formation

Summary

Although cellular reprogramming enables the generation of new cell types for disease modeling and regenerative therapies, reprogramming remains a rare cellular event. By examining reprogramming of fibroblasts into motor neurons and multiple other somatic lineages, we find that epigenetic barriers to conversion can be overcome by endowing cells with the ability to mitigate an inherent antagonism between transcription and DNA replication. We show that transcription factor overexpression induces unusually high rates of transcription and that sustaining hypertranscription and transgene expression in hyperproliferative cells early in reprogramming is critical for successful lineage conversion. However, hypertranscription impedes DNA replication and cell proliferation, processes that facilitate reprogramming. We identify a chemical and genetic cocktail that dramatically increases the number of cells capable of simultaneous hypertranscription and hyperproliferation by activating topoisomerases. Further, we show that hypertranscribing, hyperproliferating cells reprogram at 100-fold higher, near-deterministic rates. Therefore, relaxing biophysical constraints overcomes molecular barriers to cellular reprogramming.

Keywords

reprogramming
transcription factor
genomic instability
topoisomerase
single-cell RNA-seq
transcription rate
hypertranscription
p53
Repsox

Cited by (0)

5

Present address: Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

6

These authors contributed equally

7

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