Efficient prime editing in mouse brain, liver and heart with dual AAVs
Efficient prime editing in mouse brain, liver and heart with dual AAVs
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DOI：
10.1038/s41587-023-01758-z
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发表时间：
2023-05-04
影响因子：
46.9
通讯作者：
Liu, David R.
中科院分区：
文献类型：
--
作者：
Davis, Jessie R.;Banskota, Samagya;Liu, David R.
实现启动编辑对遗传疾病研究和治疗的承诺需要有效的方法来在体内传递启动编辑器（PEs）。在这里，我们描述了在体内限制腺相关病毒（AAV）介导的引物编辑的瓶颈，以及具有PE表达增加、引物编辑指导RNA稳定性和DNA修复调节的AAV-PE载体的开发。由此产生的双aav系统，v1em和v3em PE-AAV，能够在小鼠大脑（皮质效率高达42%），肝脏（高达46%）和心脏（高达11%）中进行治疗相关的先导编辑。我们应用这些系统在体内为阿尔茨海默病的星形胶质细胞和冠状动脉疾病的肝细胞安装假定的保护性突变。在体内用v3em PE-AAV进行引物编辑不会引起可检测到的脱靶效应或肝酶或组织学的显着变化。优化的PE-AAV系统支持迄今为止报道的最高未富集水平的体内引体编辑，促进了具有遗传成分的疾病的研究和潜在治疗。用AAV载体将Prime编辑器有效地传递到小鼠器官。
Realizing the promise of prime editing for the study and treatment of genetic disorders requires efficient methods for delivering prime editors (PEs) in vivo. Here we describe the identification of bottlenecks limiting adeno-associated virus (AAV)-mediated prime editing in vivo and the development of AAV-PE vectors with increased PE expression, prime editing guide RNA stability and modulation of DNA repair. The resulting dual-AAV systems, v1em and v3em PE-AAV, enable therapeutically relevant prime editing in mouse brain (up to 42% efficiency in cortex), liver (up to 46%) and heart (up to 11%). We apply these systems to install putative protective mutations in vivo for Alzheimer's disease in astrocytes and for coronary artery disease in hepatocytes. In vivo prime editing with v3em PE-AAV caused no detectable off-target effects or significant changes in liver enzymes or histology. Optimized PE-AAV systems support the highest unenriched levels of in vivo prime editing reported to date, facilitating the study and potential treatment of diseases with a genetic component.Prime editors are efficiently delivered to mouse organs with AAV vectors.