Comparison of DNA targeting CRISPR editors in human cells

人类细胞中 DNA 靶向 CRISPR 编辑器的比较

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作者:Hongxin Huang, Weiqi Lv, Jinhe Li, Guanjie Huang, Zhihong Tan, Yongfei Hu, Shufeng Ma, Xin Zhang, Linxuan Huang, Ying Lin

Background

Profiling and comparing the performance of current widely used DNA targeting CRISPR systems provide the basic information for the gene-editing toolkit and can be a useful resource for this field. In the current study, we made a parallel comparison between the recently reported miniature Cas12f1 (Un1Cas12f1 and AsCas12f1) and the widely used Cas12a and Cas9 nucleases in mammalian cells.

Conclusion

The comparison provided the editing properties of the widely used DNA-targeting CRISPR systems in the gene-editing field.

Results

We found that as a CRISPRa activator, Un1Cas12f1 could induce gene expression with a comparable level to that of Cas12a and Cas9, while as a DNA cleavage editor, Cas12f1 exhibited similar properties to Cas12a, like high specificity and dominantly induced deletions over insertions, but with less activity. In contrast, wild-type SpCas9 showed the highest activity, lowest specificity, and induced balanced deletions and insertions. Thus, Cas12f1 is recommended for gene-activation-based applications, Cas12a is for therapy applications, and wild-type Cas9 is for in vitro and animal investigations.

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