Dib Carla, Queenan Jack A, Swartzrock Leah, Willner Hana, Denis Morgane, Ahmed Nouraiz, Moulana Zada Fareha, Borges Beltran, Charlesworth Carsten T, Lum Tony, Yates Bradley P, Kwon Caleb Y, Scorzo Augustino V, Davis Scott C, Davis Jessie R, He Ran, Xie Jun, Gao Guangping, MacKenzie Tippi C, Liu David R, Newby Gregory A, Czechowicz Agnieszka D
Department of Pediatrics, Division of Hematology, Oncology, Stem Cell Transplantation and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA, USA.
Nat Commun. 2025 Jul 31;16(1):7017. doi: 10.1038/s41467-025-61449-y.
Gene editing technologies have revolutionized therapies for numerous genetic diseases. However, in vivo gene editing hinges on identifying efficient delivery vehicles for editing in targeted cell types, a significant hurdle in fully realizing its therapeutic potential. A model system to rapidly evaluate systemic gene editing would advance the field. Here, we develop the GFP-on reporter mouse, which harbors a nonsense mutation in a genomic EGFP sequence correctable by adenine base editor (ABE) among other genome editors. The GFP-on system was validated using single and dual adeno-associated virus (AAV9) encoding ABE8e and sgRNA. Intravenous administration of AAV9-ABE8e-sgRNA into adult GFP-on mice results in EGFP expression consistent with the tropism of AAV9. Intrahepatic delivery of AAV9-ABE8e-sgRNA into GFP-on fetal mice restores EGFP expression in AAV9-targeted organs lasting at least six months post-treatment. The GFP-on model provides an ideal platform for high-throughput evaluation of emerging gene editing tools and delivery modalities.
基因编辑技术已经彻底改变了多种遗传疾病的治疗方法。然而，体内基因编辑依赖于为靶向细胞类型的编辑识别高效的递送载体，这是充分实现其治疗潜力的一个重大障碍。一个能够快速评估全身基因编辑的模型系统将推动该领域的发展。在此，我们开发了绿色荧光蛋白开启（GFP-on）报告小鼠，其基因组绿色荧光蛋白（EGFP）序列中存在一个无义突变，可被腺嘌呤碱基编辑器（ABE）及其他基因组编辑器校正。使用编码ABE8e和sgRNA的单腺相关病毒（AAV9）和双腺相关病毒（AAV9）对GFP-on系统进行了验证。将AAV9-ABE8e-sgRNA静脉注射到成年GFP-on小鼠体内，导致EGFP表达与AAV9的嗜性一致。将AAV9-ABE8e-sgRNA肝内注射到GFP-on胎鼠体内，可在治疗后至少六个月内恢复AAV9靶向器官中的EGFP表达。GFP-on模型为新兴基因编辑工具和递送方式的高通量评估提供了一个理想平台。
