小柯机器人

研究利用1型CRISPR–Cas系统靶向转录调控
2019-09-24 13:32

美国杜克大学Charles A. Gersbach研究组利用1型CRISPR–Cas系统在人类细胞中实现靶向转录调控。相关论文发表2019年9月23日在线发表在《自然—生物技术》杂志上。

研究人员证明了1型CRISPR-Cas系统可以在哺乳动物细胞中表达,并用于DNA靶向和转录控制。研究人员将来自大肠杆菌和单核细胞增多性李斯特菌(Listeria monocytogenes)的1型CRISPR-Cas系统的I型变体重新利用,其通过多组分RNA引导的复合物(被称为Cascade)靶向DNA。研究人员证明了人类细胞中Cascade的表达、复合物的形成和核的定位,并证明了可编程CRISPR RNA(crRNA)介导的人类基因组中特定基因座的靶向。通过将激活和抑制结构域绑定到Cascade,研究人员调节了人类细胞中靶向内源基因的表达。

这项研究证明了Cascade可以作为一种基于CRISPR的技术来靶向真核基因调控,并凸显了1型CRISPR–Cas系统可进一步被开发。

据了解,2型CRISPR–Cas系统,例如Cas9和Cas12,已被广泛用于靶向真核基因组中的DNA序列。但是,占据了自然界中所有CRISPR系统90%的1型CRISPR-Cas系统在基因组工程应用中仍未开发。

附:英文原文

Title: Targeted transcriptional modulation with type I CRISPR–Cas systems in human cells

Author: Adrian Pickar-Oliver, Joshua B. Black, Mae M. Lewis, Kevin J. Mutchnick, Tyler S. Klann, Kylie A. Gilcrest, Madeleine J. Sitton, Christopher E. Nelson, Alejandro Barrera

Issue&Volume: 2019-09-23

Abstract: 

Class 2 CRISPR–Cas systems, such as Cas9 and Cas12, have been widely used to target DNA sequences in eukaryotic genomes. However, class 1 CRISPR–Cas systems, which represent about 90% of all CRISPR systems in nature, remain largely unexplored for genome engineering applications. Here, we show that class 1 CRISPR–Cas systems can be expressed in mammalian cells and used for DNA targeting and transcriptional control. We repurpose type I variants of class 1 CRISPR–Cas systems from Escherichia coli and Listeria monocytogenes, which target DNA via a multi-component RNA-guided complex termed Cascade. We validate Cascade expression, complex formation and nuclear localization in human cells, and demonstrate programmable CRISPR RNA (crRNA)-mediated targeting of specific loci in the human genome. By tethering activation and repression domains to Cascade, we modulate the expression of targeted endogenous genes in human cells. This study demonstrates the use of Cascade as a CRISPR-based technology for targeted eukaryotic gene regulation, highlighting class 1 CRISPR–Cas systems for further exploration.

DOI: 10.1038/s41587-019-0235-7

Source:https://www.nature.com/articles/s41587-019-0235-7

Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:68.164
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex


本期文章:《自然—生物技术》:Online/在线发表

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