Compact zinc finger architecture utilizing toxin-derived cytidine deaminases for highly efficient base editing in human cells.
Friedrich FauserBhakti N KadamSebastian Arangundy-FranklinJessica E DavisVishvesha VaidyaNicola J SchmidtGarrett LewDanny F XiaRakshaa MureliColman NgYuanyue ZhouNicholas A ScarlottJason EshlemanYuri R BendañaDavid A ShivakAndreas ReikPatrick LiGregory D DavisJeffrey C MillerPublished in: Nature communications (2024)
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 tox , that can act upon double-stranded DNA. Here, we target DddA tox 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 tox 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.
Keyphrases
- crispr cas
- genome editing
- highly efficient
- circulating tumor
- nucleic acid
- escherichia coli
- single molecule
- cell free
- endothelial cells
- induced apoptosis
- binding protein
- gene expression
- copy number
- drug delivery
- signaling pathway
- cancer therapy
- cell cycle arrest
- cell proliferation
- oxidative stress
- endoplasmic reticulum stress
- pi k akt
- genome wide analysis