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A Practical Guide for CRISPR-Cas9-Induced Mutations in Axolotls
Konstantinos Sousounis1, Katharine Courtemanche1, Jessica L Whited2,3,4
1Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 22, 2022
Summary
This guide details using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) gene editing in axolotls. Learn to modify the axolotl genome for functional gene studies and analyze the results effectively.
Area of Science:
- Developmental Biology
- Genetics
- Amphibian Research
Background:
- The axolotl (Ambystoma mexicanum) is a key model organism in regeneration research.
- Genetic manipulation is crucial for understanding axolotl development and regeneration.
- CRISPR-Cas9 technology offers precise genome editing capabilities.
Purpose of the Study:
- To provide a comprehensive guide for CRISPR-Cas9 genome editing in axolotls.
- To enable researchers to investigate gene function in axolotls.
- To facilitate the generation and analysis of gene-edited axolotl models.
Main Methods:
- Gene sequence retrieval and annotation confirmation.
- CRISPR target design and guide RNA synthesis.
- Indel analysis to confirm editing events.
- Screening methods for identifying gene-edited (Crispant) axolotls.
Main Results:
- Demonstration of a step-by-step workflow for CRISPR application in axolotls.
- Methodology for efficient identification of successful genome edits.
- Establishment of protocols for generating functional genomics data in axolotls.
Conclusions:
- CRISPR technology is a powerful and accessible tool for axolotl genome engineering.
- This guide empowers researchers to study gene function and developmental processes in axolotls.
- Genome editing in axolotls opens new avenues for regenerative biology research.
