This content is Free Access.
Chapters
- 00:04Introduction
- 00:53C. elegans Gonad Preparation and Microinjection
- 01:56Injected Worm Recovery and Transfer
- 02:37PickingC. elegans for Screening
- 03:17Single Worm Lysis and PCR
- 04:22Restriction Digestion and Agarose Gel Electrophoresis
- 05:08Identification of Positively Edited and Homozygous Edited Worms
- 06:01Results: Representative C. elegans rbm-3.2 Gene Editing and Screening
- 07:10Conclusion
The goal of this protocol is to enable seamless CRISPR/Cas9 editing of the C. elegans genome using assembled ribonucleoprotein complexes and the dpy-10 co-CRISPR marker for screening. This protocol can be used to make a variety of genetic modifications in C. elegans including insertions, deletions, gene replacements and codon substitutions.
