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Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells
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Chapters
- 00:05Title
- 00:56Transfection
- 02:40Culturing Clones
- 04:23Allele-specific Primer Design
- 08:27Genotyping the Deletion and Analyzing qPCR Results
- 09:42Analysis of CRISPR/Cas9 Mediated Deletion in F1 ES Cells
- 11:05Conclusion
Experimental validation of enhancer activity is best approached by loss-of-function analysis. Presented here is an efficient protocol that uses CRISPR/Cas9 mediated deletion to study allele-specific regulation of gene transcription in F1 ES cells which contain a hybrid genome (Mus musculus129 x Mus castaneus).
