title: RNA processing enables predictable programming of gene expression.
abstract: Complex interactions among genetic components often result in variable systemic performance in designed multigene systems. Using the bacterial clustered regularly interspaced short palindromic repeat (CRISPR) pathway we develop a synthetic RNA-processing platform, and show that efficient and specific cleavage of precursor mRNA enables reliable and predictable regulation of multigene operons. Physical separation of linked genetic elements by CRISPR-mediated cleavage is an effective strategy to achieve assembly of promoters, ribosome binding sites, cis-regulatory elements, and riboregulators into single- and multigene operons with predictable functions in bacteria. We also demonstrate that CRISPR-based RNA cleavage is effective for regulation in bacteria, archaea and eukaryotes. Programmable RNA processing using CRISPR offers a general approach for creating context-free genetic elements and can be readily used in the bottom-up construction of increasingly complex biological systems in a plug-and-play manner.
authors.affiliation: Department of Bioengineering, University of California Berkeley, Berkeley, California, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Qi
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Haurwitz
authors.forename: Wenjun
authors.initials: W
authors.lastname: Shao
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Doudna
authors.forename: Adam P
authors.initials: AP
authors.lastname: Arkin
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005818:Genetic Engineering; D055029:Inverted Repeat Sequences / Q000235:genetics; D009876:Operon / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D020506:Untranslated Regions / Q000235:genetics
chemical_list: D020506:Untranslated Regions