Compact zinc finger architecture utilizing toxin-derived cytidine deaminases for highly efficient base editing in human cells.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 38360922.
- Also identified by DOI 10.1038/s41467-024-45100-w and PMC identifier 10869815.
- 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
Nucleobase editors represent an emerging technology that enables precise single-base edits to the genomes of eukaryotic cells. Most nucleobase editors use deaminase domains that act upon single-stranded DNA and require RNA-guided proteins such as Cas9 to unwind the DNA prior to editing. However, the most recent class of base editors utilizes a deaminase domain, DddA<sub>tox</sub>, that can act upon double-stranded DNA. Here, we target DddA<sub>tox</sub> fragments and a FokI-based nickase to the human CIITA gene by fusing these domains to arrays of engineered zinc fingers (ZFs). We also identify a broad variety of Toxin-Derived Deaminases (TDDs) orthologous to DddA<sub>tox</sub> that allow us to fine-tune properties such as targeting density and specificity. TDD-derived ZF base editors enable up to 73% base editing in T cells with good cell viability and favorable specificity.
Medical subject headings
- Gene Editing
- Cytidine Deaminase