Development of a nanoparticle-based gene editing technology for neurological applications

摘要 本补充提案重点研究CRISPR Cas9核糖核蛋白(RNPs)是否与 在R6/2小鼠模型上，细胞穿透肽(CPPs)可以将小鼠从亨廷顿病中拯救出来。 这些实验是基于我们最近的研究，证明了CPP可以有效地转染CRISPR 使用对流增强递送(CED)进行颅内注射后，纹状体中的Cas9-RNPs神经元。 我们选择了2/6的小鼠模型进行这些研究，因为这个模型最近用 基于Cas9的基因组编辑，使用AAV提供的SaCas9。在这份提案中，我们将在初步的基础上 研究并将通过CED将与CPPS复合的Cas9-RNP注射到纹状体和丘脑，并将 研究我们是否可以编辑亨廷顿蛋白(HTT)基因来拯救亨廷顿病小鼠。如果成功， 这些实验将确定大脑中产生治疗结果所需的基因组编辑水平 在亨廷顿的动物模型中。

Abstract This supplement proposal focuses on investigating if CRISPR Cas9 ribonucleoproteins (RNPs) complexed to cell-penetrating peptides (CPPs) can rescue mice from Huntington’s disease, using the R6/2 mouse model. These experiments are based upon our recent studies demonstrating that CPPs can efficiently transfect CRISPR Cas9-RNPs neurons in the striatum after an intracranial injection using convection enhanced delivery (CED). We have selected the 2/6 mouse model for these studies because this model has recently been rescued with Cas9 based genome editing, using AAV delivered SaCas9. In this proposal we will build on our Preliminary Studies and will inject Cas9-RNP complexed to CPPs into the striatum and thalamus, via CED, and will investigate if we can edit the huntingtin (HTT) gene to rescue mice from Huntington’s disease. If successful, these experiments will identify the levels of genome editing needed in the brain to generate therapeutic outcomes in the Huntington’s animal model.