<br>通过 TATA 盒中单核苷酸的变化调节植物中 CaMV 35S 启动子的转录活性,ACS Synthetic Biology

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通过 TATA 盒中单核苷酸的变化调节植物中 CaMV 35S 启动子的转录活性
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2022-12-23 , DOI: 10.1021/acssynbio.2c00457
Stephanie C Amack _{1,

2} , Savio S Ferreira _{1,

2} , Mauricio S Antunes _{1,

2}

Affiliation

合成生物学使用基因编码的设备和电路在活细胞和生物体中实现新颖的复杂功能。这些遗传电路的一个标志是它们各个部分之间的相互作用，根据预先定义的规则，处理细胞信息并产生电路输出或响应。随着遗传回路中单个组件数量的增加，实现正确行为所需的相互作用数量也随之增加，因此更需要微调每个组件的表达水平。转录启动子在遗传回路中发挥着关键的调节作用，因为它们影响产生的 RNA 和蛋白质的水平。在植物等多细胞生物中，它们还可以确定基因表达的发育、空间和组织特异性模式。花椰菜花叶病毒 (CaMV 35S) 的 35S 启动子广泛应用于植物生物技术中，以指导多种植物物种的高水平基因表达。我们通过向启动子的 TATA 盒序列引入所有单核苷酸取代，产生了 CaMV 35S 启动子的 21 个变体的文库。然后，我们表征了纯合转基因植物中所有变体的活性，并表明其中一些变体的活性低于植物中的野生型。这些启动子变体可用于微调植物中合成遗传电路的行为。

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Tuning the Transcriptional Activity of the CaMV 35S Promoter in Plants by Single-Nucleotide Changes in the TATA Box

Synthetic biology uses genetically encoded devices and circuits to implement novel complex functions in living cells and organisms. A hallmark of these genetic circuits is the interaction among their individual parts, according to predefined rules, to process cellular information and produce a circuit output or response. As the number of individual components in a genetic circuit increases, so does the number of interactions needed to achieve the correct behavior, and hence, a greater need to fine-tune the levels of expression of each component. Transcriptional promoters play a key regulatory role in genetic circuits, as they influence the levels of RNA and proteins produced. In multicellular organisms, such as plants, they can also determine developmental, spatial, and tissue-specific patterns of gene expression. The 35S promoter from the Cauliflower Mosaic Virus (CaMV 35S) is widely used in plant biotechnology to direct high levels of gene expression in a variety of plant species. We produced a library of 21 variants of the CaMV 35S promoter by introducing all single nucleotide substitutions to the promoter’s TATA box sequence. We then characterized the activity of all variants in homozygous transgenic plants and showed that some of these variants have lower activity than the wild type in plants. These promoter variants could be used to fine-tune the behavior of synthetic genetic circuits in plants.

更新日期：2022-12-23

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