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利用 Piezo1 机械敏感离子通道的磁遗传学进行 CRISPR 基因编辑。

Magnetogenetics with Piezo1 Mechanosensitive Ion Channel for CRISPR Gene Editing.

机构信息

Center for Nanomedicine, Institute for Basic Science (IBS), Seoul 03722, Republic of Korea.

Department of Chemistry, Yonsei University, Seoul 03722, Republic of Korea.

出版信息

Nano Lett. 2022 Sep 28;22(18):7415-7422. doi: 10.1021/acs.nanolett.2c02314. Epub 2022 Sep 7.

Abstract

Regulation of genetic activity in single cells and tissues is pivotal to determine key cellular functions in current biomedicine, yet the conventional biochemical activators lack spatiotemporal precision due to the diffusion-mediated slow kinetics and nonselectivity. Here, we describe a magnetogenetic method for target-specific activation of a clustered regularly interspaced short palindromic repeats (CRISPR) system for the regulation of intracellular proteins. We used magnetomechanical force generated by the magnetic nanostructure to activate pre-encoded Piezo1, the mechanosensitive ion channel, on the target cell. The activated Piezo1 further triggers the intracellular Ca signaling pathway, inducing the pre-encoded genes to express genes of interest (GOIs), which is Cas9 protein for the CRISPR regulation of the target proteins. We demonstrated that this magnetogenetic CRISPR system successfully edits the target genome for both and pseudo- environments, providing a versatile magnetic platform for remote gene editing of animals with various size scales.

摘要

调控单细胞和组织内的基因活性对当前的生物医学来说至关重要,因为传统的生化激活剂由于扩散介导的缓慢动力学和非选择性而缺乏时空精度。在这里,我们描述了一种磁遗传学方法,用于针对簇状规则间隔的短回文重复序列 (CRISPR) 系统的靶特异性激活,以调节细胞内蛋白。我们使用磁纳米结构产生的磁机械力来激活目标细胞上预先编码的 Piezo1,即机械敏感离子通道。激活的 Piezo1 进一步触发细胞内 Ca 信号通路,诱导预先编码的基因表达感兴趣的基因 (GOIs),对于 CRISPR 靶蛋白调控来说是 Cas9 蛋白。我们证明了这种磁遗传学 CRISPR 系统可以成功地编辑真实和伪环境中的靶基因组,为各种大小尺度的动物提供了一种通用的远程基因编辑磁性平台。

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