Tool Preparation and Single-Cell Stage Egg Collection
1:34
Pico-Injection
3:36
Screening of Morpholino-Injected Individuals, CRISPR Mutagenesis, and Tol2 Transmission
4:35
Results: Representative Post-Pico-Injection Functional and Phenotypical Analyses
6:27
Conclusion
Transcript
Cave animals provide a compelling system for investigating diverse morphological, developmental, and behavioral traits. But one of the limiting factors in establishing this model system is a lack of genetic tools that would allow us to manipulate
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We describe approaches for the manipulation of genes in the evolutionary model system Astyanax mexicanus. Three different techniques are described: Tol2-mediated transgenesis, targeted manipulation of the genome using CRISPR/Cas9, and knockdown of expression using morpholinos. These tools should facilitate the direct investigation of genes underlying the variation between surface- and cave-dwelling forms.