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An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing
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Chapters
- 00:04Title
- 01:01Methodological Overview
- 01:54Design and Construction of gRNA Expression Vector
- 04:19Designing and Constructing the HDR Donor
- 06:01Fly Genetics and Screening for Genome Editing
- 08:13Results: Validation of Tissue-specific and Conditional bnl-LexA Expression in Different Tissues
- 09:10Conclusion
Here, we present a method for generating tissue-specific binary transcription systems in Drosophila by replacing the first coding exon of genes with transcription drivers. The CRISPR/Cas9-based method places a transactivator sequence under the endogenous regulation of a replaced gene, and consequently facilitates transctivator expression exclusively in gene-specific spatiotemporal patterns.
