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090			_aB-17820
245	1	0	_aTargeted Transcriptional Activation in Plants Using a Potent Dead Cas9-Derived Synthetic Gene Activator.
490	0		_vCurrent Protocols in Molecular Biology, 127(1), p.e89, 2019
520	3		_aGenetic tools for specific perturbation of endogenous gene expression are highly desirable for interrogation of plant gene functions and improvement of crop traits. Synthetic transcriptional activators derived from the CRISPR/Cas9 system are emerging as powerful new tools for activating the endogenous expression of genes of interest in plants. These synthetic constructs, generated by tethering transcriptional activation domains to a nuclease?dead Cas9 (dCas9), can be directed to the promoters of endogenous target genes by single guide RNAs (sgRNAs)to activate transcription. Here, we provide a detailed protocol for targeted transcriptional activation in plants using a recently developed, highly potent dCas9 gene activator construct referred to as dCas9?TV. This protocol covers selection of sgRNA targets, construction of sgRNA expression cassettes, and screening for an optimal sgRNA using a protoplast?based promoter?luciferase assay. Finally, the dCas9?TV gene activator coupled with the optimal sgRNA is delivered into plants via Agrobacterium?mediated transformation, thereby enabling robust upregulation of target gene expression in transgenic Arabidopsis and rice plants. © 2019 by John Wiley & Sons, Inc.
650	1	4	_aCRISPR
650	1	4	_aDCAS9 GENE ACTIVATOR
650	1	4	_aPLANTS
650	1	4	_aPROMOTER
650	1	4	_aTRANSCRIPTIONAL ACTIVATION
700	1	2	_aLi, Z.
700	1	2	_aWang, F.
700	1	2	_aLi, J.F.
856	4	0	_uhttps://drive.google.com/file/d/1zVYRnP4LWFwTtuzgJu8U8wt8bZMcf-nC/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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