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JoVE Journal

Genetics

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Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

9.1K Views

10:46 min

June 2nd, 2018

June 2nd, 2018

9,135 Views

0:05

Title

0:37

Guide RNA Design

3:37

Guide RNA Cloning

6:27

Transfection of Enhancer-I, Cell Harvest, and RNA Extraction

8:21

Results: Guide RNA Design/Cloning and Evaluation of Gene Expression in Response to Enhancer-i

9:23

Conclusion

Transkript

This protocol describes the steps needed to design and perform multiplexed targeting of enhancers with the deactivating fusion protein SID4X-dCas9-KRAB, also known as enhancer interference (Enhancer-i). This protocol enables the identification of enhancers that regulate gene expression and facilitates the dissection of relationships between enhancers regulating a common target gene.

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