Multiple Gene Repression in Cyanobacteria Using CRISPRi
Multiple Gene Repression in Cyanobacteria Using CRISPRi
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DOI：
10.1021/acssynbio.5b00264
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发表时间：
2016-03-01
影响因子：
4.7
通讯作者：
Hudson, Elton P.
中科院分区：
文献类型：
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作者：
Yao, Lun;Cengic, Ivana;Hudson, Elton P.
我们描述了成簇规则间隔短回文重复干扰（CRISPRi）在蓝细菌集胞藻属PCC 6803模型中基因抑制的应用。来自II型的核酸酶缺陷型Cas9。使用化脓性链球菌的CRISPR/Cas将绿色荧光蛋白（GFP）抑制至可忽略的水平。CRISPRi还用于在氮饥饿期间抑制碳储存化合物聚羟基丁酸酯（PHB）和糖原的形成。作为CRISPRi在基础和应用蓝藻研究中的潜力的一个例子，我们同时敲低了4种推定的醛还原酶和脱氢酶，抑制率为50-95%。这项工作还表明，紧密抑制的启动子允许蓝藻中的诱导型和可逆的CRISPRi。
We describe the application of clustered regularly interspaced short palindromic repeats interference (CRISPRi) for gene repression in the model cyanobacterium Synechcocystis sp. PCC 6803. The nuclease-deficient Cas9 from the type-II. CRISPR/Cas of Streptrococcus pyogenes was used to repress green fluorescent protein (GFP) to negligible levels. CRISPRi was also used to repress formation of carbon storage compounds polyhydroxybutryate (PHB) and glycogen during nitrogen starvation. As an example of the potential of CRISPRi for basic and applied cyanobacteria research, we simultaneously knocked down 4 putative aldehyde reductases and dehydrogenases at 50-95% repression. This work also demonstrates that tightly repressed promoters allow for inducible and reversible CRISPRi in cyanobacteria.