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多重基因编辑减少了原发性超氧沙流症1型中氧酸盐的产生

Rui Zheng¹, De-Xin Zhang¹, Yan-Jiao Shao¹

¹Department of Pediatric Urology, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China.

Zoological research

|September 27, 2023

概括

使用CRISPR-Cpf1 (来自Prevotella和Francisella1的集群定期间隔的短Palindromic重复) 的多重基因编辑,在患有初级超氧沙流症I型 (PH1) 的老鼠中,通过准Hao1和Ldha基因,成功降低了氧酸盐生产和脏损伤.

科学领域:

生物化学生物化学
遗传学是一个遗传学.
分子生物学分子生物学

背景情况:

一次性高氧化尿症I型 (PH1) 是一种严重的遗传疾病,其特征是过度的氧酸盐生产,导致损伤和衰竭.
目前PH1的治疗策略有限,需要新的方法来减少氧沙酸盐合成.

研究的目的:

调查使用CRISPR-Cpf1系统多重基因编辑方法的有效性,以在PH1.1的老鼠模型中同时准肝氧酸氧化酶1 (Hao1) 和乳酸脱酶A (Ldha) 基因.
评估该策略在减少氧沙酸盐形成和改善PH1相关病理方面的潜力.

主要方法:

选CRISPRRNA (crRNA) 对,以老鼠Hao1和Ldha基因为目标,以获得最佳的效率和特异性.
在体内向PH1大鼠输送腺相关病毒 (AAV) - AsCpf1,以实现Hao1和Ldha的多重基因组编辑.
评估基因组编辑效率,基因表达变化,尿液中氧酸盐水平,损伤和甲.

主要成果:

在体内实现了对Hao1和Ldha基因的高效基因组编辑,主要是通过小的删除,导致基因表达减少.
在PH1大鼠中,用AAV-AsCpf1治疗显著降低了尿氧酸盐水平,并减轻了损伤和甲.
没有发现显著的肝毒性,肝外基因组编辑或非目标效应.

结论:

AAV-AsCpf1系统展示了强大的多重基因编辑能力,可以同时准参与氧酸盐合成的多个基因.
这项研究为基于多重基因组编辑的基因疗法提供了概念验证,作为PH1的有希望的治疗策略.
这种方法为开发新型治疗方法提供了潜在的途径,通过解决其根本原因来管理或治愈PH1.

关键词:

基因组编辑基因组编辑氧化酸氧化酶 1 的含量过高氧化尿症是什么?乳酸脱酶的使用方法

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