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Efficient DNA base editing via an optimized DYW-like deaminase

Kweon, J.; Park, S.; Jeon, M. Y.; Lim, K.; Jang, G.; Jang, A.-H.; Lee, M.; Seok, C.; Lee, C.; Park, S.; Ahn, J.; Jang, J.; Sung, Y.; Kim, D.; Kim, Y.

2024-05-16 molecular biology
10.1101/2024.05.15.594452 bioRxiv
Show abstract

CRISPR-based cytosine base editors enable precise genome editing without inducing double-stranded DNA breaks, yet traditionally depend on a limited selection of deaminases from the APOBEC/AID or TadA families. Here, we introduce SsCBE, a novel CRISPR-based cytosine base editor utilizing SsdAtox, a DYW-like deaminase derived from the toxin of Pseudomonas syringae. Strategic engineering of SsdAtox has led to remarkable improvements in the base editing efficiency (by up to 8.4-fold) and specificity for SsCBE, while concurrently reducing cytotoxicity. Exhibiting exceptional versatility, SsCBE was delivered and efficiently applied using diverse delivery methods, including the engineered virus-like particles (eVLPs). Its application has enabled targeted cytosine base editing in mouse zygotes and pioneering edits in mitochondrial DNA. The advent of SsCBE marks a significant advancement in the CRISPR toolkit, providing a versatile tool for advanced research and therapeutic strategies.

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