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Nat Biotechnol. 2012 Oct;30(10):1002-6. doi: 10.1038/nbt.2355. Epub 2012 Sep 16.

RNA processing enables predictable programming of gene expression.

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1
Department of Bioengineering, University of California Berkeley, Berkeley, California, USA.

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.

PMID:
22983090
DOI:
10.1038/nbt.2355
[Indexed for MEDLINE]
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