Generation and Analysis of CCM Phenotypes in C. elegans

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

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.

Keywords: C. elegans; CRISPR; Cerebral cavernous malformations; Drug screen; Forward genetic screen; High-throughput screen; Suppressor screen; ccm-2; ccm-3; kri-1.

MeSH terms

  • Alleles
  • Animals
  • CRISPR-Cas Systems
  • Caenorhabditis elegans
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism
  • Gene Editing
  • Genetic Predisposition to Disease
  • Hemangioma, Cavernous, Central Nervous System / diagnosis
  • Hemangioma, Cavernous, Central Nervous System / etiology*
  • Hemangioma, Cavernous, Central Nervous System / metabolism*
  • Mutagenesis
  • Mutation
  • Phenotype*
  • Ribonucleoproteins / metabolism

Substances

  • Caenorhabditis elegans Proteins
  • Ribonucleoproteins