Engineered miniature CRISPR-Cas system for mammalian genome regulation and editing

• DOI: 10.1016/j.molcel.2021.08.008
• Publication Date: 2021-09-03T14:35:26Z
• Authors: Xiaoshu Xu; Augustine Chempar...; Leiping Zeng; Hannah R. Kempton; Stephen Shang; Muneaki Nakamura; Lei S. Qi
• Keywords: Genome Engineering; Synthetic Biology; Guide RNA

Compact and versatile CRISPR-Cas systems will enable genome engineering applications through high-efficiency delivery in a wide variety of contexts. Here, we create an efficient miniature Cas system (CasMINI) engineered from the type V-F Cas12f (Cas14) by guide RNA protein engineering, which is less than half size currently used CRISPR (Cas9 or Cas12a). We demonstrate that CasMINI can drive high levels gene activation (up to thousands-fold increases), while natural fails function mammalian cells. show has comparable activities Cas12a for activation, highly specific, allows robust base editing editing. expect be broadly useful cell therapy ex vivo vivo.