Engineering cell signaling using tunable CRISPR–Cpf1-based transcription factors

Yuchen Liu (), Jinghong Han, Zhicong Chen, Hanwei Wu, Hongsong Dong and Guohui Nie ()
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Yuchen Liu: The First Affiliated Hospital of Shenzhen University
Jinghong Han: The First Affiliated Hospital of Shenzhen University
Zhicong Chen: The First Affiliated Hospital of Shenzhen University
Hanwei Wu: The First Affiliated Hospital of Shenzhen University
Hongsong Dong: The First Affiliated Hospital of Shenzhen University
Guohui Nie: The First Affiliated Hospital of Shenzhen University

Nature Communications, 2017, vol. 8, issue 1, 1-8

Abstract: Abstract The catalytically dead Cpf1 endonuclease from Acidaminococcus sp. BV3L6 (dAsCpf1) has been used to construct effective transcriptional repressors in bacteria and plants. However, it is still unclear if dAsCpf1 can function in human cells as a transcriptional regulator or a signal conductor. Here, we repurpose the dAsCpf1 system in human cells for a variety of functions, including the activation or repression of gene transcription. Moreover, we construct programmable ligand-controlled dAsCpf1 systems either by coupling crRNAs with engineered riboswitches or by fusing dAsCpf1 proteins with G protein-coupled receptors. These generalizable approaches allow us to regulate the transcription of endogenous genes in response to diverse classes of ligands, thus constructing artificial signaling pathways with rewired cellular input–output behaviors. The systems exhibit signal amplification, an important feature in cell signaling, when multiple crRNAs are processed from a single transcript. The results provide a robust and efficient platform for engineering customized cell signaling circuits.

Date: 2017
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DOI: 10.1038/s41467-017-02265-x

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