2. Academic Validation
  3. Spatially concentrated adenine base editors efficiently correct PLP1 mutations in oligodendrocytes

Spatially concentrated adenine base editors efficiently correct PLP1 mutations in oligodendrocytes

  • Nucleic Acids Res. 2026 Feb 24;54(5):gkag156. doi: 10.1093/nar/gkag156.
Chi Zhang 1,  Ke Ye 1,  Yafang Shang 2,  Yixuan Song 1,  Pingping Li 1,  Xinyue Jiang 1,  Cuiping Yang 3,  Aibin Liang 4,  Jian Zhang 5,  Feilong Meng 2,  Mingliang Zhang 1 3 6 7
Affiliations

Affiliations

  • 1 Department of Histoembryology, Genetics and Developmental Biology, Shanghai Key Laboratory of Reproductive Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
  • 2 Key Laboratory of RNA Innovation, Science and Engineering, Shanghai Academy of Natural Sciences (SANS), Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
  • 3 The International Peace Maternity and Child Health Hospital, Shanghai Key Laboratory of Embryo Original Diseases, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China.
  • 4 Department of Hematology, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China.
  • 5 Key Laboratory of Protection, Development and Utilization of Medicinal Resources in Liupanshan Area, Ministry of Education, Peptide & Protein Drug Research Center, College of Pharmacy, Ningxia Medical University, Yinchuan, Ningxia 750004, China.
  • 6 Department of Histology and Embryology, Zunyi Medical University, Zunyi, Guizhou 563000, China.
  • 7 School of Basic Medical Sciences, Ningxia Medical University, Yinchuan, Ningxia 750004, China.
PMID: 41736546 DOI: 10.1093/nar/gkag156
Abstract

Oligodendrocytes (OLs), the myelinating cells of the central nervous system, are particularly prone to pathogenic G-to-A mutations, such as PLP1A243V, which causes Pelizaeus-Merzbacher disease (PMD), a lethal hypomyelinating disorder lacking effective therapy. Although adenine base editors (ABEs) can in principle correct such mutations, their application in OLs is limited by inefficient on-target editing. Here, we develop a spatially concentrated ABE (cABE) strategy that enhances editing by promoting nuclear translocation of tRNA Adenosine Deaminase (TadA*) from the cytoplasm. Using a SunTag-based multivalent recruitment system, TadA* is locally enriched at genomic targets (cABE-1.0), achieving robust editing in vitro. To enable in vivo delivery while preserving high efficiency and fidelity, SpCas9 is replaced with compact eNme2-C Cas9, generating an AAV-compatible variant (cABE-2.0). Notably, cABE-2.0 forms dynamic nuclear puncta with properties of liquid-liquid phase separation, enhancing on-target editing while substantially reducing transcriptome-wide RNA off-target effects. Functionally, cABE-2.0 efficiently corrects the PLP1A243V mutation in OLs, restores Plp subcellular localization, and rescues myelination-related phenotypes. These findings demonstrate that spatial reorganization, rather than increasing intrinsic catalytic activity of TadA*, provides a distinct principle for improving base editing in difficult-to-edit cell types, such as OLs, offering a mechanistic and technical framework for gene therapy of PMD and related myelin disorders.

Figures
Products