Department of Bioengineering, University of California Berkeley, Berkeley, California, USA.
Nat Biotechnol. 2012 Oct;30(10):1002-6. doi: 10.1038/nbt.2355. Epub 2012 Sep 16.
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.
在设计多基因系统时，遗传成分之间的复杂相互作用通常会导致系统表现出现可变性。我们利用细菌成簇规律间隔短回文重复序列（CRISPR）途径开发了一种合成 RNA 处理平台，并表明前体 mRNA 的有效和特异性切割能够可靠且可预测地调节多基因操纵子。通过 CRISPR 介导的切割将连锁遗传元件物理分离是一种有效的策略，可将启动子、核糖体结合位点、顺式调控元件和核糖开关组装到单基因和多基因操纵子中，从而在细菌中实现可预测的功能。我们还证明，基于 CRISPR 的 RNA 切割可有效调节细菌、古菌和真核生物。使用 CRISPR 进行可编程 RNA 处理为创建无上下文遗传元件提供了一种通用方法，并且可以以即插即用的方式方便地用于以自下而上的方式构建越来越复杂的生物系统。
