Enhancing CRISPR-Cas for disease modeling in Xenopus

项目总结 CRISPR-CAS系统的最新发展改变了许多细胞的非遗传模型 生物学，然而其基因组编辑能力的充分应用在很大程度上被限制在温血动物身上 易受当前CRISPR-Cas介导的基因方法影响的物种和基因 颠覆。生活在较低温度下的几个物种，如非洲爪哇和海洋模型Lolio 是模拟人类疾病的理想系统，但目前精确的基因组编辑方法 这些物种是不够的。此外，由于它们的基因组结构，许多高度保守的 致病基因对于目前的基因组编辑方法来说是难以实现的。我们建议扩大 CRISPR-CAS介导的基因组编辑工具包在人类疾病建模领域的应用 两个方面：1)能够影响靶基因的低温CRISPR-Cas系统的建立 插入在较低温度下茁壮成长的模式物种，以及2)开发一种新的靶向系统 应用CRISPR-Cas文库进行基因破坏。

PROJECT SUMMARY The recent development of the CRISPR-Cas system has transformed many non-genetic models of cellular biology, yet the full application of its genome editing capabilities has been largely restricted to warm-blooded species and to genes which are easily amenable to current methods of CRISPR-Cas mediated gene disruption. Several species which live at lower temperatures, such as Xenopus and the marine models Loligo and Limulus, are ideal systems to model human disease, yet current methods of precise genome editing in these species are inadequate. Furthermore, due to their genomic architecture, many highly-conserved disease-causing genes are refractory to current methods of genome editing. We propose to expand the CRISPR-Cas mediated genome editing toolkit in the field of human disease modeling by driving innovation in two areas: 1) the development of a low-temperature CRISPR-Cas system capable of effecting targeted gene insertion in model species which thrive at lower temperatures, and 2) to develop a nascent system of targeted gene disruption by the application of CRISPR-Cas libraries.