Genes & Human Disease Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA.
Institute of Modern Aquaculture Science and Engineering, School of Life Sciences, South China Normal University, Guangzhou, Guangdong, 510631, China.
Nat Commun. 2024 Jul 4;15(1):5613. doi: 10.1038/s41467-024-49943-1.
Advancements in CRISPR technology, particularly the development of base editors, revolutionize genetic variant research. When combined with model organisms like zebrafish, base editors significantly accelerate and refine in vivo analysis of genetic variations. However, base editors are restricted by protospacer adjacent motif (PAM) sequences and specific editing windows, hindering their applicability to a broad spectrum of genetic variants. Additionally, base editors can introduce unintended mutations and often exhibit reduced efficiency in living organisms compared to cultured cell lines. Here, we engineer a suite of adenine base editors (ABEs) called ABE-Ultramax (Umax), demonstrating high editing efficiency and low rates of insertions and deletions (indels) in zebrafish. The ABE-Umax suite of editors includes ABEs with shifted, narrowed, or broadened editing windows, reduced bystander mutation frequency, and highly flexible PAM sequence requirements. These advancements have the potential to address previous challenges in disease modeling and advance gene therapy applications.
CRISPR 技术的进步，特别是碱基编辑器的发展，彻底改变了遗传变异研究。当与斑马鱼等模式生物结合使用时，碱基编辑器可显著加速和优化遗传变异的体内分析。然而，碱基编辑器受到原间隔基序（PAM）序列和特定编辑窗口的限制，限制了其在广泛遗传变异中的应用。此外，碱基编辑器可能会引入意外突变，并且与培养细胞系相比，其在活体生物中的效率往往较低。在这里，我们设计了一套腺嘌呤碱基编辑器（ABEs），称为 ABE-Ultramax（Umax），在斑马鱼中表现出高效的编辑效率和较低的插入和缺失（indels）率。ABE-Umax 编辑器套件包括编辑窗口移位、变窄或变宽、旁反应突变频率降低以及高度灵活的 PAM 序列要求的 ABE。这些进展有可能解决疾病建模中的先前挑战，并推进基因治疗应用。
