Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, 91125, USA.
Department of Bioengineering and Imperial College Centre for Synthetic Biology, Imperial College London, London, UK.
Nat Commun. 2020 Mar 9;11(1):1281. doi: 10.1038/s41467-020-15053-x.
Multiplexed CRISPR technologies, in which numerous gRNAs or Cas enzymes are expressed at once, have facilitated powerful biological engineering applications, vastly enhancing the scope and efficiencies of genetic editing and transcriptional regulation. In this review, we discuss multiplexed CRISPR technologies and describe methods for the assembly, expression and processing of synthetic guide RNA arrays in vivo. Applications that benefit from multiplexed CRISPR technologies, including cellular recorders, genetic circuits, biosensors, combinatorial genetic perturbations, large-scale genome engineering and the rewiring of metabolic pathways, are highlighted. We also offer a glimpse of emerging challenges and emphasize experimental considerations for future studies.
多重 CRISPR 技术，其中大量的 gRNA 或 Cas 酶同时表达，极大地促进了遗传编辑和转录调控的范围和效率，为强大的生物工程应用提供了便利。在这篇综述中，我们讨论了多重 CRISPR 技术，并描述了在体内组装、表达和处理合成向导 RNA 阵列的方法。受益于多重 CRISPR 技术的应用，包括细胞记录器、遗传电路、生物传感器、组合遗传扰动、大规模基因组工程和代谢途径的重新布线，都得到了强调。我们还展望了新兴的挑战，并强调了未来研究的实验考虑因素。
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