Targeted genome regulation via synthetic programmable transcriptional regulators.
Regulation of gene transcription controls cellular functions and coordinates responses to developmental, physiological and environmental cues. Precise and efficient molecular tools are needed to characterize the functions of single and multiple genes in linear and interacting pathways in a native context. Modular DNA-binding domains from zinc fingers (ZFs) and transcriptional activator-like proteins (TALE) are amenable to bioengineering to bind DNA target sequences of interest. As a result, ZF and TALE proteins were used to develop synthetic programmable transcription factors. However, these systems are limited by the requirement to re-engineer proteins for each new target sequence. The clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR associated 9 (Cas9) genome editing tool was recently repurposed for targeted transcriptional regulation by inactivation of the nuclease activity of Cas9. Due to the facile engineering, simplicity, precision and amenability to library construction, the CRISPR/Cas9 system is poised to revolutionize the functional genomics field across diverse eukaryotic species. In this review, we discuss the development of synthetic customizable transcriptional regulators and provide insights into their current and potential applications, with special emphasis on plant systems, in characterization of gene functions, elucidation of molecular mechanisms and their biotechnological applications.
a Laboratory for Genome Engineering, Center for Desert Agriculture and Division of Biological and Environmental Sciences and Engineering , King Abdullah University of Science and Technology , Thuwal , Jeddah , Saudi Arabia.
Agnieszka
A
Piatek
a Laboratory for Genome Engineering, Center for Desert Agriculture and Division of Biological and Environmental Sciences and Engineering , King Abdullah University of Science and Technology , Thuwal , Jeddah , Saudi Arabia.
Magdy M
MM
Mahfouz
D001709:Biotechnology / Q000379:methods* / Q000639:trends; D064113:CRISPR-Cas Systems / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D000072669:Gene Editing / Q000379:methods* / Q000639:trends; D005818:Genetic Engineering / Q000379:methods* / Q000639:trends; D016678:Genome; D000072417:Protein Domains / Q000235:genetics; D014158:Transcription, Genetic; D015533:Transcriptional Activation / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics
D004268:DNA-Binding Proteins
CRISPR/Cas9 system; functional genomics; synthetic site-specific nucleases; synthetic transcriptional regulators; targeted genome regulation; transcriptional activator-like effectors; zinc fingers