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生成CRISPR / Cas9媒介1アレル欠失マウス胚性幹細胞でエンハンサー機能を研究するために、
JoVE Journal
Biologia do Desenvolvimento
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JoVE Journal Biologia do Desenvolvimento
Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells
DOI:

11:31 min

April 02, 2016

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Capítulos

  • 00:05Título
  • 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

Summary

Tadução automática

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).

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