title: CRISPR-Cas type II-based Synthetic Biology applications in eukaryotic cells.
abstract: The CRISPR-Cas system has rapidly reached a huge popularity as a new, powerful method for precise DNA editing and genome reengineering. In Synthetic Biology, the CRISPR-Cas type II system has inspired the construction of a novel class of RNA-based transcription factors. In their simplest form, they are made of a CRISPR RNA molecule, which targets a promoter sequence, and a deficient Cas9 (i.e. deprived of any nuclease activity) that has been fused to an activation or a repression domain. Up- and downregulation of single genes in mammalian and yeast cells have been achieved with satisfactory results. Moreover, the construction of CRISPR-based transcription factors is much simpler than the assembly of synthetic proteins such as the Transcription Activator-Like effectors. However, the feasibility of complex synthetic networks fully based on the CRISPR-dCas9 technology has still to be proved and new designs, which take into account different CRISPR types, shall be investigated.
authors.affiliation: a School of Life Science and Technology, Harbin Institute of Technology , Harbin , P.R. China.
authors.forename: Mario Andrea
authors.identifier: 0000-0002-5102-1069
authors.initials: MA
authors.lastname: Marchisio
authors.affiliation: a School of Life Science and Technology, Harbin Institute of Technology , Harbin , P.R. China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Huang
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D005057:Eukaryotic Cells; D005786:Gene Expression Regulation; D006801:Humans; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000378:metabolism; D058615:Synthetic Biology / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D014157:Transcription Factors
keywords: CRISPR; Cas9; Synthetic Biology; gene circuits; guide RNA