当前位置: X-MOL 学术Cell › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors.
Cell ( IF 45.5 ) Pub Date : 2018-Apr-19 , DOI: 10.1016/j.cell.2018.02.033
Silvana Konermann 1 , Peter Lotfy 1 , Nicholas J Brideau 1 , Jennifer Oki 1 , Maxim N Shokhirev 2 , Patrick D Hsu 1
Affiliation  

Class 2 CRISPR-Cas systems endow microbes with diverse mechanisms for adaptive immunity. Here, we analyzed prokaryotic genome and metagenome sequences to identify an uncharacterized family of RNA-guided, RNA-targeting CRISPR systems that we classify as type VI-D. Biochemical characterization and protein engineering of seven distinct orthologs generated a ribonuclease effector derived from Ruminococcus flavefaciens XPD3002 (CasRx) with robust activity in human cells. CasRx-mediated knockdown exhibits high efficiency and specificity relative to RNA interference across diverse endogenous transcripts. As one of the most compact single-effector Cas enzymes, CasRx can also be flexibly packaged into adeno-associated virus. We target virally encoded, catalytically inactive CasRx to cis elements of pre-mRNA to manipulate alternative splicing, alleviating dysregulated tau isoform ratios in a neuronal model of frontotemporal dementia. Our results present CasRx as a programmable RNA-binding module for efficient targeting of cellular RNA, enabling a general platform for transcriptome engineering and future therapeutic development.

中文翻译:


使用 RNA 靶向 VI-D 型 CRISPR 效应器进行转录组工程。



2 类 CRISPR-Cas 系统赋予微生物多种适应性免疫机制。在这里,我们分析了原核基因组和宏基因组序列,以确定一个未表征的 RNA 引导、RNA 靶向 CRISPR 系统家族,我们将其归类为 VI-D 型。七种不同直系同源物的生化表征和蛋白质工程产生了源自黄瘤胃球菌 XPD3002 (CasRx) 的核糖核酸酶效应子,在人类细胞中具有强大的活性。相对于不同内源转录本的 RNA 干扰,CasRx 介导的敲低表现出高效率和特异性。作为最紧凑的单效应Cas酶之一,CasRx还可以灵活地包装到腺相关病毒中。我们将病毒编码的、催化失活的 CasRx 靶向前 mRNA 的顺式元件,以操纵选择性剪接,减轻额颞叶痴呆神经元模型中 tau 亚型比例失调的情况。我们的结果表明 CasRx 作为一种可编程的 RNA 结合模块,可有效靶向细胞 RNA,为转录组工程和未来治疗开发提供通用平台。
更新日期:2018-04-26
down
wechat
bug