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Engineered minimal type I CRISPR-Cas system for transcriptional activation and base editing in human cells.

Jing GuoLuyao GongHaiying YuMing LiQiaohui AnZhenquan LiuShuru FanChangjialian YangDahe ZhaoJing HanHua Xiang
Published in: Nature communications (2024)
Type I CRISPR-Cas systems are widespread and have exhibited high versatility and efficiency in genome editing and gene regulation in prokaryotes. However, due to the multi-subunit composition and large size, their application in eukaryotes has not been thoroughly investigated. Here, we demonstrate that the type I-F2 Cascade, the most compact among type I systems, with a total gene size smaller than that of SpCas9, can be developed for transcriptional activation in human cells. The efficiency of the engineered I-F2 tool can match or surpass that of dCas9. Additionally, we create a base editor using the I-F2 Cascade, which induces a considerably wide editing window (~30 nt) with a bimodal distribution. It can expand targetable sites, which is useful for disrupting functional sequences and genetic screening. This research underscores the application of compact type I systems in eukaryotes, particularly in the development of a base editor with a wide editing window.
Keyphrases
  • crispr cas
  • genome editing
  • genome wide
  • gene expression
  • transcription factor
  • copy number
  • heat shock
  • low cost
  • mass spectrometry
  • high resolution
  • atomic force microscopy
  • genome wide analysis