Department of Histoembryology, Genetics and Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Shanghai Key Laboratory of Reproductive Medicine, Shanghai 200025, China.
Nucleic Acids Res. 2022 Sep 23;50(17):9632-9646. doi: 10.1093/nar/gkac742.
Adenine base editors (ABEs) catalyze A-to-G conversions, offering therapeutic options to treat the major class of human pathogenic single nucleotide polymorphisms (SNPs). However, robust and precise editing at diverse genome loci remains challenging. Here, using high-throughput chemical screening, we identified and validated SB505124, a selective ALK5 inhibitor, as an ABE activator. Treating cells with SB505124 enhanced on-target editing at multiple genome loci, including epigenetically refractory regions, and showed little effect on off-target conversion on the genome. Furthermore, SB505124 facilitated the editing of disease-associated genes in vitro and in vivo. Intriguingly, SB505124 served as a specific activator by selectively promoting ABE activity. Mechanistically, SB505124 promotes ABE editing, at least in part, by enhancing ABE expression and modulating DNA repair-associated genes. Our findings reveal the role of the canonical transforming growth factor-β pathway in gene editing and equip ABEs with precise chemical control.
腺嘌呤碱基编辑器（ABEs）催化 A 到 G 的转换，为治疗人类主要致病单核苷酸多态性（SNPs）提供了治疗选择。然而，在不同基因组位点实现强大且精确的编辑仍然具有挑战性。在这里，我们通过高通量化学筛选，鉴定并验证了 SB505124，一种选择性的 ALK5 抑制剂，是 ABE 的激活剂。用 SB505124 处理细胞可增强多个基因组位点的靶向编辑，包括表观遗传难治区域，并且对基因组上的非靶向转换几乎没有影响。此外，SB505124 促进了体外和体内疾病相关基因的编辑。有趣的是，SB505124 通过选择性地促进 ABE 活性来充当特异性激活剂。在机制上，SB505124 通过增强 ABE 的表达和调节与 DNA 修复相关的基因，至少部分促进了 ABE 的编辑。我们的研究结果揭示了经典转化生长因子-β 途径在基因编辑中的作用，并为 ABE 提供了精确的化学控制。
