CRISPR‐Act3.0‐Based Highly Efficient Multiplexed Gene Activation in Plants
CRISPR‐Act3.0‐Based Highly Efficient Multiplexed Gene Activation in Plants
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DOI：
10.1002/cpz1.365
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发表时间：
2022-02
期刊：
影响因子：
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通讯作者：
Changtian Pan;Yiping Qi
中科院分区：
文献类型：
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作者：
Changtian Pan;Yiping Qi
CRISPR/Cas（成簇规律间隔的短回文重复序列/CRISPR相关蛋白）介导的基因组编辑彻底改变了基础研究和植物育种。除了基因编辑之外，CRISPR/Cas 系统已被重新用作可编程转录调控平台。催化失活 Cas 变体 (dCas) 与转录激活域融合时，可以特异性激活基因组中的任何靶基因，而不会诱导 DNA 双链断裂。 CRISPR 激活能够同时激活多个基因，在识别基因调控网络和重新连接代谢途径方面具有广阔的前景。在这里，我们描述了一个基于 CRISPR-Act3.0 系统构建 dCas9 介导的多重基因激活系统的简单协议。所得载体在水稻原生质体中进行测试。 © 2022 Wiley 期刊有限责任公司。
CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR‐associated protein)‐mediated genome editing has revolutionized fundamental research and plant breeding. Beyond gene editing, CRISPR/Cas systems have been repurposed as a platform for programmable transcriptional regulation. Catalytically inactive Cas variants (dCas), when fused with transcriptional activation domains, allow for specific activation of any target gene in the genome without inducing DNA double‐strand breaks. CRISPR activation enables simultaneous activation of multiple genes, holding great promise in the identification of gene regulatory networks and rewiring of metabolic pathways. Here, we describe a simple protocol for constructing a dCas9‐mediated multiplexed gene activation system based on the CRISPR‐Act3.0 system. The resulting vectors are tested in rice protoplasts. © 2022 Wiley Periodicals LLC.