RNA-guided transcriptional regulation in planta via synthetic dCas9-based transcription factors

Agnieszka Anna Piatek, Zahir Ali, Hatoon Baazim, Lixin Li, Aala A. Abulfaraj, Sahar Alshareef, Mustapha Aouida, Magdy M. Mahfouz

Research output: Contribution to journalArticlepeer-review

182 Scopus citations

Abstract

Targeted genomic regulation is a powerful approach to accelerate trait discovery and development in agricultural biotechnology. Bacteria and archaea use clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated (Cas) regulatory systems for adaptive molecular immunity against foreign nucleic acids introduced by invading phages and conjugative plasmids. The type II CRISPR/Cas system has been adapted for genome editing in many cell types and organisms. A recent study used the catalytically inactive Cas9 (dCas9) protein combined with guide-RNAs (gRNAs) as a DNA-targeting platform to modulate gene expression in bacterial, yeast, and human cells. Here, we modified this DNA-targeting platform for targeted transcriptional regulation in planta by developing chimeric dCas9-based transcriptional activators and repressors. To generate transcriptional activators, we fused the dCas9 C-terminus with the activation domains of EDLL and TAL effectors. To generate a transcriptional repressor, we fused the dCas9 C-terminus with the SRDX repression domain. Our data demonstrate that dCas9 fusion with the EDLL activation domain (dCas9:EDLL) and the TAL activation domain (dCas9:TAD), guided by gRNAs complementary to selected promoter elements, induce strong transcriptional activation on Bs3
Original languageEnglish (US)
Pages (from-to)578-589
Number of pages12
JournalPlant Biotechnology Journal
Volume13
Issue number4
DOIs
StatePublished - Nov 14 2014

ASJC Scopus subject areas

  • Plant Science
  • Agronomy and Crop Science
  • Biotechnology

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