Zhang Xuhua, Xu Linping, Fan Ruihua, Gao Quanli, Song Yunfeng, Lyu Xiaodong, Ren Jiangtao, Song Yongping
1School of Life Sciences, Zhengzhou University, Zhengzhou, China.
2Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital, Zhengzhou, China.
Cell Discov. 2018 Jul 10;4:36. doi: 10.1038/s41421-018-0035-0. eCollection 2018.
Cpf1, an RNA-guided DNA endonuclease that belongs to a new class II CRISPR system, has recently been harnessed for genome editing. Herein, we report an RNase-resistant caged truncated pre-tRNA-like crRNA (catRNA) that confers precise and efficient gene editing with the Cpf1 (LbCpf1) and enables the reprogramming of catalytically dead LbCpf1 (dCpf1) lacking DNA endonuclease activity into a transcriptional modulator. Specific gene knock-outs and knock-ins were increased 3.2-fold and 4.3-fold, respectively, with catRNA compared to that induced by conventional crRNA. A much higher augmentation of gene disruption (up to 37-fold) was observed when electroporation was used. We report herein that catRNA enables efficient gene activation with dCpf1 activators. Our study reveals the potential of catRNA and a versatile application of the CRISPR/Cpf1 system, establishing a simple approach for selective gene perturbation in mammalian cells.
Cpf1是一种属于新型II类CRISPR系统的RNA引导的DNA内切核酸酶，最近已被用于基因组编辑。在此，我们报告了一种抗核糖核酸酶的笼化截短前体tRNA样crRNA（catRNA），它能与Cpf1（LbCpf1）实现精确高效的基因编辑，并能将缺乏DNA内切核酸酶活性的催化失活LbCpf1（dCpf1）重编程为转录调节因子。与传统crRNA诱导的相比，catRNA介导的特异性基因敲除和敲入分别增加了3.2倍和4.3倍。当使用电穿孔时，观察到基因破坏的增强幅度更高（高达37倍）。我们在此报告，catRNA能够与dCpf1激活剂实现高效的基因激活。我们的研究揭示了catRNA的潜力以及CRISPR/Cpf1系统的广泛应用，建立了一种在哺乳动物细胞中进行选择性基因干扰的简单方法。
