Dose-dependent activation of gene expression is achieved using CRISPR and small molecules that recruit endogenous chromatin machinery

Anna M. Chiarella, Kyle V. Butler, Berkley E. Gryder, Dongbo Lu, Tiffany A. Wang, Xufen Yu, Silvia Pomella, Javed Khan, Jian Jin, Nathaniel A. Hathaway

Research output: Contribution to journalLetter

Abstract

Gene expression can be activated or suppressed using CRISPR­–Cas9 systems. However, tools that enable dose-dependent activation of gene expression without the use of exogenous transcription regulatory proteins are lacking. Here we describe chemical epigenetic modifiers (CEMs) designed to activate the expression of target genes by recruiting components of the endogenous chromatin-activating machinery, eliminating the need for exogenous transcriptional activators. The system has two parts: catalytically inactive Cas9 (dCas9) in complex with FK506-binding protein (FKBP) and a CEM consisting of FK506 linked to a molecule that interacts with cellular epigenetic machinery. We show that CEMs upregulate gene expression at target endogenous loci up to 20-fold or more depending on the gene. We also demonstrate dose-dependent control of transcriptional activation, function across multiple diverse genes, reversibility of CEM activity and specificity of our best-in-class CEM across the genome.

Original languageEnglish
Pages (from-to)50-55
Number of pages6
JournalNature Biotechnology
Volume38
Issue number1
DOIs
Publication statusE-pub ahead of print - 2019

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ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Cite this

Chiarella, A. M., Butler, K. V., Gryder, B. E., Lu, D., Wang, T. A., Yu, X., Pomella, S., Khan, J., Jin, J., & Hathaway, N. A. (2019). Dose-dependent activation of gene expression is achieved using CRISPR and small molecules that recruit endogenous chromatin machinery. Nature Biotechnology, 38(1), 50-55. https://doi.org/10.1038/s41587-019-0296-7