RNA Therapeutics Institute; Program in Molecular Medicine
Amino Acids, Peptides, and Proteins | Computational Biology | Genomics | Molecular Biology | Nucleic Acids, Nucleotides, and Nucleosides
In Caenorhabditis elegans, targeted genome editing techniques are now routinely used to generate germline edits. The remarkable ease of C. elegans germline editing is attributed to the syncytial nature of the pachytene ovary which is easily accessed by microinjection. This protocol describes the step-by-step details and troubleshooting tips for the entire CRISPR-Cas genome editing procedure, including gRNA design and microinjection of ribonucleoprotein complexes, followed by screening and genotyping in C. elegans, to help accessing this powerful genetic animal system. For complete details on the use and execution of this protocol, please refer to Ghanta and Mello (2020).
CRISPR, genetics, model organisms, molecular biology
Rights and Permissions
Copyright 2021. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
DOI of Published Version
Ghanta KS, Ishidate T, Mello CC. Microinjection for precision genome editing in Caenorhabditis elegans. STAR Protoc. 2021 Sep 1;2(3):100748. doi: 10.1016/j.xpro.2021.100748. PMID: 34505086; PMCID: PMC8417391. Link to article on publisher's site
Ghanta KS, Ishidate T, Mello CC. (2021). Microinjection for precision genome editing in Caenorhabditis elegans. Open Access Publications by UMass Chan Authors. https://doi.org/10.1016/j.xpro.2021.100748. Retrieved from https://escholarship.umassmed.edu/oapubs/4978
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.