The Department of Cellular and Integrative Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA.
GenEdit Inc., Berkeley, CA, USA.
Nat Biomed Eng. 2018 Jul;2(7):497-507. doi: 10.1038/s41551-018-0252-8. Epub 2018 Jun 25.
Technologies that can safely edit genes in the brains of adult animals may revolutionize the treatment of neurological diseases and the understanding of brain function. Here, we demonstrate that intracranial injection of CRISPR-Gold, a nonviral delivery vehicle for the CRISPR-Cas9 ribonucleoprotein, can edit genes in the brains of adult mice in multiple mouse models. CRISPR-Gold can deliver both Cas9 and Cpf1 ribonucleoproteins, and can edit all of the major cell types in the brain, including neurons, astrocytes and microglia, with undetectable levels of toxicity at the doses used. We also show that CRISPR-Gold designed to target the metabotropic glutamate receptor 5 (mGluR5) gene can efficiently reduce local mGluR5 levels in the striatum after an intracranial injection. The effect can also rescue mice from the exaggerated repetitive behaviours caused by fragile X syndrome, a common single-gene form of autism spectrum disorders. CRISPR-Gold may significantly accelerate the development of brain-targeted therapeutics and enable the rapid development of focal brain-knockout animal models.
能够安全编辑成年动物大脑基因的技术可能会彻底改变神经疾病的治疗方法和对大脑功能的理解。在这里，我们证明了 CRISPR-Gold（一种用于 CRISPR-Cas9 核糖核蛋白的非病毒递送载体）的脑内注射可以在多种小鼠模型中编辑成年小鼠大脑中的基因。CRISPR-Gold 可以递送 Cas9 和 Cpf1 核糖核蛋白，并且可以编辑大脑中的所有主要细胞类型，包括神经元、星形胶质细胞和小胶质细胞，而且在使用的剂量下没有可检测到的毒性。我们还表明，针对代谢型谷氨酸受体 5（mGluR5）基因设计的 CRISPR-Gold 可以在脑内注射后有效降低纹状体局部的 mGluR5 水平。这种效果还可以挽救因脆性 X 综合征（一种常见的自闭症谱系障碍的单基因形式）而导致过度重复行为的小鼠。CRISPR-Gold 可能会显著加速针对大脑的治疗药物的开发，并能够快速开发出针对特定脑区的基因敲除动物模型。
