Engineered Transformer Base Editor with Enhanced Editing Efficiency
TL;DR
The engineered tBE (etBE) exhibited substantially enhanced editing efficiencies compared with the parental tBE, while maintaining high editing fidelity and background levels of OT mutations, and represents a highly efficient and specific base editing platform with enormous potential for both basic research and clinical applications.
Abstract
ABSTRACT Canonical cytosine base editors (CBEs) achieve precise C‐to‐T conversions without inducing DNA double‐strand breaks (DSBs), yet their clinical potential remains hampered by substantial off‐target (OT) mutations. The recently developed transformer base editor (tBE) significantly reduces both genomic and transcriptomic OT mutations by using a cleavable deoxycytidine deaminase inhibitor (dCDI) domain. However, the modest base editing efficiency limits its broader applications. Here, through rational deaminase engineering and fusion of a uracil DNA glycosylase inhibitor (UGI) domain, we developed the engineered tBE (etBE). The etBE exhibited substantially enhanced editing efficiencies compared with the parental tBE (up to 35.11‐fold improvement), while maintaining high editing fidelity and background levels of OT mutations. As a therapeutic proof‐of‐concept, dual adeno‐associated virus (AAV)‐mediated delivery of etBE targeting proprotein convertase subtilisin/kexin type 9 (PCSK9), a well‐established therapeutic target for cardiovascular diseases, was evaluated in a humanized mouse model. The treatment achieved efficient in vivo base editing (up to 35.13%), resulting in substantial reductions in plasma PCSK9 protein (24%) and low‐density lipoprotein cholesterol (LDL‐C) levels (33%), while inducing only minimal OT mutations. Collectively, etBE represents a highly efficient and specific base editing platform with enormous potential for both basic research and clinical applications.
