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  • Engineering cell signaling using tunable CRISPR-Cpf1-based transcription factors.

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

Nature communications (2017-12-14)
Yuchen Liu, Jinghong Han, Zhicong Chen, Hanwei Wu, Hongsong Dong, Guohui Nie
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Histone Deacetylase 1 (HDAC1) antibody, Mouse monoclonal, clone HDAC1-21, purified from hybridoma cell culture
Sigma-Aldrich
Anti-AsCas12a (Cpf1) antibody, Mouse monoclonal, clone AsCpf-11, purified from hybridoma cell culture