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

Link to Article in PubMed

PubMed ID

34505086

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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