Chapter title |
Multiplexed Transcriptional Activation or Repression in Plants Using CRISPR-dCas9-Based Systems
|
---|---|
Chapter number | 12 |
Book title |
Plant Gene Regulatory Networks
|
Published in |
Methods in molecular biology, June 2017
|
DOI | 10.1007/978-1-4939-7125-1_12 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7124-4, 978-1-4939-7125-1
|
Authors |
Lowder, Levi G., Paul, Joseph W., Qi, Yiping, Levi G. Lowder, Joseph W. Paul III, Yiping Qi, Joseph W. Paul |
Editors |
Kerstin Kaufmann, Bernd Mueller-Roeber |
Abstract |
Novel tools and methods for regulating in vivo plant gene expression are quickly gaining popularity and utility due to recent advances in CRISPR-dCas9 chimeric effector regulators, otherwise known as CRISPR artificial transcription factors (CRISPR-ATFs). These tools are especially useful for studying gene function and interaction within various regulatory networks. First generation CRISPR-ATFs are nuclease-deactivated (dCas9) CRISPR systems where dCas9 proteins are fused to known transcriptional activator domains (VP64) or repressor domains (SRDX). When multiple chimeric dCas9-effector fusions are guided to gene regulatory regions via CRISPR gRNAs, they can modulate expression of transcript levels in planta. The protocol presented here provides a detailed procedure for activating AtPAP1 and repressing AtCSTF64 in Arabidopsis thaliana. This protocol makes use of our plant CRISPR toolbox to streamline the assembly and cloning of multiplex CRISPR-Cas9 transcriptional regulatory constructs. |
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