Repurposing type I-F CRISPR-Cas system as a transcriptional activation tool in human cells.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 32561716.
- Also identified by DOI 10.1038/s41467-020-16880-8 and PMC identifier 7305327.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Class 2 CRISPR-Cas proteins have been widely developed as genome editing and transcriptional regulating tools. Class 1 type I CRISPR-Cas constitutes ~60% of all the CRISPR-Cas systems. However, only type I-B and I-E systems have been used to control mammalian gene expression and for genome editing. Here we demonstrate the feasibility of using type I-F system to regulate human gene expression. By fusing transcription activation domain to Pseudomonas aeruginosa type I-F Cas proteins, we activate gene transcription in human cells. In most cases, type I-F system is more efficient than other CRISPR-based systems. Transcription activation is enhanced by elongating the crRNA. In addition, we achieve multiplexed gene activation with a crRNA array. Furthermore, type I-F system activates target genes specifically without off-target transcription activation. These data demonstrate the robustness and programmability of type I-F CRISPR-Cas in human cells.
Medical subject headings
- Bacterial Proteins
- CRISPR-Associated Proteins
- CRISPR-Cas Systems
- Clustered Regularly Interspaced Short Palindromic Repeats
- Gene Editing