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CRISPR-Act2.0: An Improved Multiplexed System for Plant Transcriptional Activation

  • Aimee Malzahn
  • Yong Zhang
  • Yiping Qi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1917)

Abstract

CRISPR systems have greatly promoted research in genome editing and transcriptional regulation. CRISPR-based transcriptional repression and activation systems will be valuable for applications in engineering plant immunity, boosting metabolic production, and enhancing our knowledge of gene regulatory networks. Multiplexing of CRISPR allows multiple genes to be targeted without significant additional effort. Here, we describe our CRISPR-Act2.0 system which is an improved multiplexing transcriptional activation system in plants.

Key words

CRISPR dCas9-VP64 MCP-VP64 Plant transcriptional activation Artificial transcription factor Multiplex Golden Gate assembly Gateway cloning 

Notes

Acknowledgments

This work is supported by the NSF Plant Genome ECA-PGR Award (#1758745) of the University of Maryland, College Park, MD.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Plant Science and Landscape ArchitectureUniversity of Maryland College ParkCollege ParkUSA
  2. 2.Department of Biotechnology, School of Life Science and Technology, Center for Informational BiologyUniversity of Electronic Science and Technology of ChinaChengduChina
  3. 3.Institute for Bioscience and Biotechnology ResearchUniversity of MarylandRockvilleUSA

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