Novel methods to improve nuclease mediated homologous recombination, Administrative suppl

项目摘要 在这份行政补助金申请中，我们要求资金购买一个新的细胞， 分选仪（S3 e ™细胞分选仪，Bio-Rad）。这种最先进的细胞分选仪将使我们能够评估大的 大小基因敲入事件，包括中靶敲入、中靶插入/缺失（indel） 速率以及潜在的脱靶敲入和插入缺失事件。在Aim中 1，我们计划使用该细胞分选仪来评估mi-spCas 9-v2。在目标2中，我们将使用这种细胞分选仪 来评估mi-saCas 9在目标3中，我们将使用该细胞分选仪来评估有效性和安全性 miCas 9在临床相关应用中的概况。结果，这种新的细胞分选机，一旦 配备，将产生更有说服力的数据，以证明所提出的优势 miCas 9基因编辑核酸酶。

Project Abstract In this Administrative Supplement grant application, we request funds to purchase a new cell sorter (S3e™ Cell Sorter, Bio-Rad). This state of art cell sorter will enable us to evaluate large size gene knock-in events, including both on-target knock-in, on-target insertion/deletion (indel) rates as well as potential off-target knock-in and indel events, at the single cell resolution. In Aim 1, we plan to use this cell sorter to evaluate mi-spCas9-v2. In Aim 2, we will use this cell sorter to evalute mi-saCas9. In Aim 3, we will use this cell sorter to evaluate the efficacy and safety profiles of miCas9 in clinically relevant applications. As a results, this new cell sorter, once equipped, will generate more convincing data to demonstrate the advantages of the proposed miCas9 gene editing nucleases.