In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway
In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway
复制标题
体内基因组编辑通过 Cas9/RecA 介导的同源定向修复途径挽救光感受器变性
DOI：
10.1126/sciadv.aav3335
复制
发表时间：
2019-04-01
期刊：
影响因子：
13.6
通讯作者：
Xue, Tian
中科院分区：
文献类型：
--
作者：
Cai, Yuan;Cheng, Tianlin;Xue, Tian
尽管cas9介导的基因组编辑已被广泛用于人类遗传性疾病动物模型中的等位基因工程，但很少有基于同源定向修复（HDR）的基因编辑系统已在出生后小鼠模型中建立，以实现有效和持久的表型拯救。在这里，我们开发了一种基于HDR的Cas9/RecA系统，以精确纠正Pde6b突变，提高出生后无视网膜（rd1）小鼠的HDR效率，rd1是一种以光感受器变性和视力丧失为特征的视网膜色素变性（RP）突变模型。Cas9/RecA系统结合Cas9核酸内切酶产生双链断裂（DSBs）和细菌重组酶A （RecA）增加同源重组。我们的数据显示，Cas9/RecA处理显著促进了杆状和锥状光感受器的存活，恢复了杆状光感受器中PDE6B的表达，增强了rd1小鼠的视觉功能。因此，本研究为RP和其他遗传疾病提供了精确的治疗策略。
Although Cas9-mediated genome editing has been widely used to engineer alleles in animal models of human inherited diseases, very few homology-directed repair (HDR)-based genetic editing systems have been established in postnatal mouse models for effective and lasting phenotypic rescue. Here, we developed an HDR-based Cas9/RecA system to precisely correct Pde6b mutation with increased HDR efficiency in postnatal rodless (rd1) mice, a retinitis pigmentosa (RP) mutant model characterized by photoreceptor degeneration and loss of vision. The Cas9/RecA system incorporated Cas9 endonuclease enzyme to generate double-strand breaks (DSBs) and bacterial recombinase A (RecA) to increase homologous recombination. Our data revealed that Cas9/RecA treatment significantly promoted the survival of both rod and cone photoreceptors, restored the expression of PDE6B in rod photoreceptors, and enhanced the visual functions of rd1 mice. Thus, this study provides a precise therapeutic strategy for RP and other genetic diseases.