Dual Activation of CD95/CXCR4 in HEK293T cells by Abscisic Acid- and Gibberellin-inducible dCas9s (Transient Transfection)
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.
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Posted 25 Oct, 2016
Dual Activation of CD95/CXCR4 in HEK293T cells by Abscisic Acid- and Gibberellin-inducible dCas9s (Transient Transfection)
Posted 25 Oct, 2016
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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