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Chapters
- 00:04Introduction
- 00:58Preparation of CRISPR Gene Editing and Guide RNA Design to Generate miRNA Cluster Knockout Cell Lines
- 02:06Generation of Stable Lentiviral Cell Lines Carrying the DOX-Inducible Cas9 Expression Cassette
- 03:20Performing CRISPR Reactions by Cas9 Induction and Synthetic Guide RNA Transfection of Cells Using a High-Throughput Format
- 07:22PCR Genotyping of CRISPR Cell Lines Using Crude Cell Lysates
- 08:37Results: High-Throughput CRISPR Experimental Design to Generate Multiple miR-888 Cluster Deletion Combinations
- 09:51Conclusion
This protocol describes a high-throughput clustered regularly interspaced short palindromic repeats (CRISPR) gene editing workflow for microRNA cluster network analysis that allows the rapid generation of a panel of genetically modified cell lines carrying unique miRNA cluster member deletion combinations as large as 35 kb within a single experiment.
