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bioRxiv · 10.1101/253757

Phosphorothioate-modified DNA oligonucleotides inactivate CRISPR-Cpf1 mediated genome editing

Li, B.·Zeng, C.·Li, W.·Zhang, X.·Luo, X.·Zhao, W.·Zhang, C.·Dong, Y.
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Abstract

CRISPR-Cpf1, a microbial adaptive immune system discovered from Prevotella and Francisella 1, employs a single-stranded CRISPR RNA (crRNA) to induce double stranded DNA breaks1. To modulate genome editing activity of Cpf1 in human cells, we designed a series of crRNA variants including DNA-crRNA and RNA-crRNA duplexes, and identified that phosphorothioate (PS)-modified DNA-crRNA duplex completely blocked the function of Cpf1 mediated gene editing. More importantly, without prehybridization, this PS-modified DNA was able to regulate Cpf1 activity in a time-and dose-dependent manner. Mechanistic studies indicate that PS-modified DNA oligonucleotides hinder the binding between Cpf1-crRNA complex and target DNA substrate. Consequently, phosphorothioate-modified DNA oligonucleotides provide a tunable platform to inactivate Cpf1 mediated genome editing.

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Li, B., Zeng, C., Li, W., Zhang, X., Luo, X., Zhao, W., Zhang, C., Dong, Y.. 2018-02-06. Phosphorothioate-modified DNA oligonucleotides inactivate CRISPR-Cpf1 mediated genome editing. https://doi.org/10.1101/253757

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