Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells

Sakthi D. Moorthy; Jennifer A. Mitchell

doi:10.3791/53552

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Generazione CRISPR / Cas9 Mediated monoallelic eliminazioni per studiare la funzione Enhancer nel topo cellule staminali embrionali

Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells

JoVE Journal
Biologia dello sviluppo

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JoVE Journal Biologia dello sviluppo

Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells

Generazione CRISPR / Cas9 Mediated monoallelic eliminazioni per studiare la funzione Enhancer nel topo cellule staminali embrionali

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14,145 Views

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11:31 min

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April 02, 2016

DOI:

10.3791/53552-v

DOI:

10.3791/53552-v

•

11:31 min

April 02, 2016

•

14126 Views

Sakthi D. Moorthy¹, Jennifer A. Mitchell¹

¹Department of Cell and Systems Biology,University of Toronto

Summary

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).