瑞士巴塞尔大学Randall J. Platt研究组的一项最新工作实现了在单个转录本上编码Cas12a和CRISPR矩阵，从而对基因组多个位点进行编辑。这一研究成果发表在2019年9月出版的国际学术期刊《自然—方法学》上。

研究人员发现，Cas12a和成簇规则间隔短回文重复（CRISPR）矩阵可以通过添加一个稳定的三级RNA结构实现在单个转录本中进行编码。通过利用该系统，研究人员使用在单个质粒上传递的多达25个单独的CRISPR RNA来验证了针对内源性靶标可实现的复杂基因组工程技术（包括组成型、条件性、诱导型、正交和多位点）。这个方法提供了一个强大的平台来研究和调控复杂细胞行为的复杂遗传程序。

据研究人员介绍，同时修改多个遗传元件的能力将有助于阐明和控制复杂细胞功能背后的基因相互作用和网络。然而，当前的基因组工程技术在可以同时进行操控的数量和类型方面受到限制。

附：英文原文

Title: Multiplexed genome engineering by Cas12a and CRISPR arrays encoded on single transcripts

Author: Carlo C. Campa, Niels R. Weisbach, Antnio J. Santinha, Danny Incarnato, Randall J. Platt

Issue&Volume: Volume 16 Issue 9

Abstract: The ability to modify multiple genetic elements simultaneously would help to elucidate and control the gene interactions and networks underlying complex cellular functions. However, current genome engineering technologies are limited in both the number and the type of perturbations that can be performed simultaneously. Here, we demonstrate that both Cas12a and a clustered regularly interspaced short palindromic repeat (CRISPR) array can be encoded in a single transcript by adding a stabilizer tertiary RNA structure. By leveraging this system, we illustrate constitutive, conditional, inducible, orthogonal and multiplexed genome engineering of endogenous targets using up to 25 individual CRISPR RNAs delivered on a single plasmid. Our method provides a powerful platform to investigate and orchestrate the sophisticated genetic programs underlying complex cell behaviors.

DOI: 10.1038/s41592-019-0508-6

Source:https://www.nature.com/articles/s41592-019-0508-6