Cell & Bioscience (Jan 2023)

Comparison of DNA targeting CRISPR editors in human cells

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

DOI
https://doi.org/10.1186/s13578-023-00958-z
Journal volume & issue
Vol. 13, no. 1
pp. 1 – 10

Abstract

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

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