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Method Article
Lei S Qi
Stanford University
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
The use of dCas9-based transcriptional activators and repressors facilitates manipulation of endogenous gene expression. However, precise temporal control of their activities and independent control of discrete gene targets in the same cell remain difficult. Here, we describe a method for utilizing orthogonal and small-molecule inducible dCas9-based transcriptional regulators for independently activating expression of CD95 and CXCR4 in HEK293T cells via transient transfection. This protocol can be easily adapted for different gene targets by using alternative sgRNAs, providing a customizable platform for flexible gene regulation.
Keywords
CRISPR, Cas9, Gene regulation, Synthetic biology, Inducible
Figure 1
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The authors declare no competing financial interests.
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Molecular Biology
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Method Article
Lei S Qi
Stanford University
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 25 Oct, 2016
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Molecular Biology
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
The use of dCas9-based transcriptional activators and repressors facilitates manipulation of endogenous gene expression. However, precise temporal control of their activities and independent control of discrete gene targets in the same cell remain difficult. Here, we describe a method for utilizing orthogonal and small-molecule inducible dCas9-based transcriptional regulators for independently activating expression of CD95 and CXCR4 in HEK293T cells via transient transfection. This protocol can be easily adapted for different gene targets by using alternative sgRNAs, providing a customizable platform for flexible gene regulation.
Figure 1
Figure 2
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