在秀丽隐杆线虫中生成和分析 CCM 表型。

Generation and Analysis of CCM Phenotypes in C. elegans.

机构信息

Developmental and Stem Cell Biology Program, The Hospital for Sick Children, Peter Gilgan Centre for Research and Learning, Toronto, ON, Canada.

Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.

出版信息

Methods Mol Biol. 2020;2152:191-205. doi: 10.1007/978-1-0716-0640-7_15.

DOI:10.1007/978-1-0716-0640-7_15

PMID:32524554

Abstract

This chapter presents methods for exploiting the powerful tools available in the nematode worm Caenorhabditis elegans to understand the in vivo functions of cerebral cavernous malformation (CCM) genes and the organization of their associated signaling pathways. Included are methods for assessing phenotypes caused by loss-of-function mutations in the worm CCM genes kri-1 and ccm-3, CRISPR-based gene editing techniques, and protocols for conducting high-throughput forward genetic and small molecule screens.

摘要

本章介绍了利用秀丽隐杆线虫这一强大工具的方法，以了解脑动静脉畸形 (CCM) 基因的体内功能及其相关信号通路的组成。其中包括评估秀丽隐杆线虫 CCM 基因 kri-1 和 ccm-3 功能缺失突变引起的表型的方法、基于 CRISPR 的基因编辑技术，以及进行高通量正向遗传学和小分子筛选的方案。