Efficient precise in vivo base editing in adult dystrophic mice.
Efficient precise in vivo base editing in adult dystrophic mice.
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DOI：
10.1038/s41467-021-23996-y
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发表时间：
2021-06-17
影响因子：
16.6
通讯作者：
Han R
中科院分区：
文献类型：
--
作者：
Xu L;Zhang C;Li H;Wang P;Gao Y;Mokadam NA;Ma J;Arnold WD;Han R
碱基编辑的最新进展为精确纠正致病突变创造了令人兴奋的机会。然而，碱基编辑器的大尺寸及其遗传的脱靶活性对体内碱基编辑提出了挑战。此外，突变位点附近的原型间隔区邻近基序（PAM）的要求进一步限制了靶向可行性。在这里，我们修改了NG靶向腺嘌呤碱基编辑器（iABE-NGA），以克服这些挑战，并证明了精确编辑成年小鼠杜氏肌营养不良症（DMD）突变的高效率。AAV 9-iABE-NGA的全身递送导致肌营养不良蛋白恢复和功能改善。在AAV 9-iABE-NGA处理后10个月，在mdx 4cv小鼠心脏中测量到肌营养不良蛋白的几乎完全拯救，在骨骼肌纤维中具有高达15%的拯救。脱靶活性保持较低，未检测到明显毒性。这项研究强调了使用iABE-NGA进行永久碱基编辑治疗单基因疾病的前景。碱基编辑是一种用于校正导致杜氏肌营养不良症（DMD）的突变的方法，但限制在于需要特定的PAM基序和超过腺相关病毒（AAV）包装能力的大尺寸。在这里，作者修改了NG靶向腺嘌呤碱基编辑器以识别更广泛的PAM，设计了一种内含肽分裂策略以将否则过大的腺嘌呤碱基编辑器包装到AAV中，并显示当全身注射到DMD小鼠模型中时，它有效地恢复了肌肉和心脏中的肌营养不良蛋白表达
Recent advances in base editing have created an exciting opportunity to precisely correct disease-causing mutations. However, the large size of base editors and their inherited off-target activities pose challenges for in vivo base editing. Moreover, the requirement of a protospacer adjacent motif (PAM) nearby the mutation site further limits the targeting feasibility. Here we modify the NG-targeting adenine base editor (iABE-NGA) to overcome these challenges and demonstrate the high efficiency to precisely edit a Duchenne muscular dystrophy (DMD) mutation in adult mice. Systemic delivery of AAV9-iABE-NGA results in dystrophin restoration and functional improvement. At 10 months after AAV9-iABE-NGA treatment, a near complete rescue of dystrophin is measured in mdx4cv mouse hearts with up to 15% rescue in skeletal muscle fibers. The off-target activities remains low and no obvious toxicity is detected. This study highlights the promise of permanent base editing using iABE-NGA for the treatment of monogenic diseases. Base editing is one approach used to correct mutations causing cause Duchenne muscular dystrophy (DMD), but limitations are in the requirement for a specific PAM motif and the large size beyond the packaging capacity of adeno-associated virus (AAV). Here, the authors modify the NG-targeting adenine base editor to recognize a broader PAM, devise an intein split strategy to package the otherwise oversized adenine base editor into AAV, and show it efficiently restores dystrophin expression in muscle and heart when systemically injected in a mouse model of DMD
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DOI：
10.1073/pnas.86.4.1292
发表时间：
1989-02-01
影响因子：
11.1
作者：
CHAPMAN, VM;MILLER, DR;CASKEY, CT
通讯作者：
CASKEY, CT
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
20.1
作者：
El Refaey M;Xu L;Gao Y;Canan BD;Adesanya TMA;Warner SC;Akagi K;Symer DE;Mohler PJ;Ma J;Janssen PML;Han R
通讯作者：
Han R
影响因子：
56.9
作者：
Jin, Shuai;Zong, Yuan;Gao, Caixia
通讯作者：
Gao, Caixia
影响因子：
46.9
作者：
Gaudelli, Nicole M.;Lam, Dieter K.;Ciaramella, Giuseppe
通讯作者：
Ciaramella, Giuseppe