UMMS Affiliation
RNA Therapeutics Institute; Program in Molecular Medicine
Publication Date
2021-09-01
Document Type
Article
Disciplines
Amino Acids, Peptides, and Proteins | Computational Biology | Genomics | Molecular Biology | Nucleic Acids, Nucleotides, and Nucleosides
Abstract
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).
Keywords
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
10.1016/j.xpro.2021.100748
Source
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
Journal/Book/Conference Title
STAR protocols
Related Resources
PubMed ID
34505086
Repository Citation
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.
Included in
Amino Acids, Peptides, and Proteins Commons, Computational Biology Commons, Genomics Commons, Molecular Biology Commons, Nucleic Acids, Nucleotides, and Nucleosides Commons