Cpf1, a type-V CRISPR–Cas effector endonuclease, exhibits gene-editing activity in human cells through a single RNA-guided approach. Here, we report the design and assessment of an array of 42 types of engineered Acidaminococcus sp. Cpf1 (AsCpf1) CRISPR RNA (crRNA) and 5 types of AsCpf1 mRNA in human cell lines. We show that the top-performing modified crRNA (cr3′5F, containing five 2′-fluoro ribose at the 3′ terminus) and AsCpf1 mRNA (full ψ-modification) improved gene-cutting efficiency by, respectively, 127% and 177%, with respect to unmodified crRNA and plasmid-encoding AsCpf1. We also show that the combination of cr3′5F and ψ-modified AsCpf1 or Lachnospiraceae bacterium Cpf1 (LbCpf1) mRNA augmented gene-cutting efficiency by over 300% with respect to the same control, and discovered that 11 out of 16 crRNAs from Cpf1 orthologues enabled genome editing in the presence of AsCpf1. Engineered CRISPR–Cpf1 systems should facilitate a broad range of genome editing applications.

中文翻译：

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工程CRISPR–Cpf1 crRNA和mRNA最大化基因组编辑效率

Cpf1是一种V型CRISPR–Cas效应核酸内切酶，通过一种RNA指导的方法在人类细胞中表现出基因编辑活性。在这里，我们报告设计和评估的42种类型的工程酸氨基球菌sp的数组。人细胞系中的Cpf1（AsCpf1）CRISPR RNA（crRNA）和5种类型的AsCpf1 mRNA。我们显示，性能最高的修饰crRNA（cr3'5F，在3'末端包含五个2'-氟核糖）和AsCpf1 mRNA（完全ψ-修饰）分别将基因切割效率提高了127％和177％关于未修饰的crRNA和编码质粒的AsCpf1。我们还表明，cr3'5F和ψ修饰的AsCpf1或湖螺科细菌的组合Cpf1（LbCpf1）mRNA与同一个对照相比，将基因切割效率提高了300％以上，并且发现在Cpf1直向同源物中16个crRNA中有11个在AsCpf1存在的情况下能够进行基因组编辑。CRISPR-Cpf1工程系统应促进广泛的基因组编辑应用。