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Capitoli
- 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
Denne protokol beskriver en high-throughput clustered regelmæssigt interspaced short palindromic repeats (CRISPR) genredigeringsarbejdsgang til mikroRNA-klyngenetværksanalyse, der muliggør hurtig generering af et panel af genetisk modificerede cellelinjer, der bærer unikke miRNA-klyngemedlemssletningskombinationer så store som 35 kb inden for et enkelt eksperiment.
