Enhancer RNAs pp 235-250 | Cite as

Targeted Gene Activation Using RNA-Guided Nucleases

  • Alexander Brown
  • Wendy S. Woods
  • Pablo Perez-PineraEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1468)


The discovery of the prokaryotic CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated) system and its adaptation for targeted manipulation of DNA in diverse species has revolutionized the field of genome engineering. In particular, the fusion of catalytically inactive Cas9 to any number of transcriptional activator domains has resulted in an array of easily customizable synthetic transcription factors that are capable of achieving robust, specific, and tunable activation of target gene expression within a wide variety of tissues and cells. This chapter describes key experimental design considerations, methods for plasmid construction, gene delivery protocols, and procedures for analysis of targeted gene activation in mammalian cell lines using CRISPR-Cas transcription factors.

Key words

Genome engineering Synthetic biology RNA-guided nucleases CRISPR-Cas9 Gene expression Gene activation Transcription 


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Alexander Brown
    • 1
  • Wendy S. Woods
    • 1
  • Pablo Perez-Pinera
    • 1
    Email author
  1. 1.Department of BioengineeringUniversity of Illinois at Urbana-ChampaignUrbanaUSA

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