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

Handle URI:
http://hdl.handle.net/10754/563859
Title:
RNA-guided transcriptional regulation in planta via synthetic dCas9-based transcription factors
Authors:
Piatek, Agnieszka Anna ( 0000-0001-9587-6360 ) ; Ali, Zahir ( 0000-0002-7814-8908 ) ; Baazim, Hatoon; Li, Lixin; Abulfaraj, Aala Abdulaziz Hussien; Alshareef, Sahar ( 0000-0002-0377-1113 ) ; Aouida, Mustapha ( 0000-0001-7945-5320 ) ; Mahfouz, Magdy M. ( 0000-0002-0616-6365 )
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
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division; Bioscience Program; Center for Desert Agriculture
Publisher:
Wiley-Blackwell
Journal:
Plant Biotechnology Journal
Issue Date:
14-Nov-2014
DOI:
10.1111/pbi.12284
Type:
Article
ISSN:
14677644
Sponsors:
This study is funded by King Abdullah University of Science and Technology (KAUST). The authors thank Lei S. Qi for providing the Cas9 and dCas9 plasmid constructs. The authors would like to thank Moussa Benhamed and Axel de Julien de Zelicourt for their helpful comments on qPCR analysis. Moreover, authors thank the anonymous reviewers for their valuable comments which helped improve the manuscript. No conflict of interest declared.
Appears in Collections:
Articles; Bioscience Program; Center for Desert Agriculture; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorPiatek, Agnieszka Annaen
dc.contributor.authorAli, Zahiren
dc.contributor.authorBaazim, Hatoonen
dc.contributor.authorLi, Lixinen
dc.contributor.authorAbulfaraj, Aala Abdulaziz Hussienen
dc.contributor.authorAlshareef, Saharen
dc.contributor.authorAouida, Mustaphaen
dc.contributor.authorMahfouz, Magdy M.en
dc.date.accessioned2015-08-03T12:17:17Zen
dc.date.available2015-08-03T12:17:17Zen
dc.date.issued2014-11-14en
dc.identifier.issn14677644en
dc.identifier.doi10.1111/pbi.12284en
dc.identifier.urihttp://hdl.handle.net/10754/563859en
dc.description.abstractTargeted 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 Bs3en
dc.description.sponsorshipThis study is funded by King Abdullah University of Science and Technology (KAUST). The authors thank Lei S. Qi for providing the Cas9 and dCas9 plasmid constructs. The authors would like to thank Moussa Benhamed and Axel de Julien de Zelicourt for their helpful comments on qPCR analysis. Moreover, authors thank the anonymous reviewers for their valuable comments which helped improve the manuscript. No conflict of interest declared.en
dc.publisherWiley-Blackwellen
dc.subjectChimeric dCas9 transcriptional activators and repressorsen
dc.subjectEDLL activation domainen
dc.subjectSRDX repressor domainen
dc.subjectSynthetic transcriptional regulatorsen
dc.subjectTargeted genomic regulationen
dc.titleRNA-guided transcriptional regulation in planta via synthetic dCas9-based transcription factorsen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.contributor.departmentBioscience Programen
dc.contributor.departmentCenter for Desert Agricultureen
dc.identifier.journalPlant Biotechnology Journalen
kaust.authorAli, Zahiren
kaust.authorBaazim, Hatoonen
kaust.authorLi, Lixinen
kaust.authorAouida, Mustaphaen
kaust.authorMahfouz, Magdy M.en
kaust.authorPiatek, Agnieszka Annaen
kaust.authorAbulfaraj, Aala Abdulaziz Hussienen
kaust.authorAlshareef, Saharen
All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.