Sense and sensibility: of synthetic biology and the redesign of bioreporter circuits.
Sense and sensibility: of synthetic biology and the redesign of bioreporter circuits.
复制标题
DOI：
10.1111/1751-7915.13955
复制
发表时间：
2022-01
影响因子：
5.7
通讯作者：
Wang B
中科院分区：
文献类型：
--
作者：
Belkin S;Wang B
人们很容易推测，六十年前，当 Jacob 和 Monod 提出他们的 lac 操纵子模型时（Jacob 和 Monod，1961），他们已经看到了 lacZ 基因的未来，它不仅可以编码二糖切割器，也可以作为美丽且开创性的基因调控模型的组成部分，而且还可以作为基因激活的通用报告者。事实上，报告基因技术迅速成为研究基因表达调控的基本工具。然而，几十年后，同样的方法才首次报道了一种经过基因工程改造的微生物，可以对环境污染物进行准确、特异和灵敏的分析（King 等，1990）。 “全细胞生物传感器”一词很快投入使用，并伴随着一些语义争议：纯粹主义者将“生物传感器”一词视为一种硬件设备，其中生物实体（例如酶、抗体、寡核苷酸或活细胞）作为其传感组件（IUPAC，2017）；根据这种观点，微生物菌株，尽管其重新设计的复杂性，可以被称为“传感器菌株”或“生物报告器”，但绝不是“生物传感器”。然而，早在这场语言争论成为问题之前，Sayler 小组的一篇开创性文章（King 等人，1990）描述了一种基于假单胞菌的生物发光萘传感器。继第一个基于大肠杆菌的汞传感器（Selifonova 等人，1993）发布之后，很快又出现了许多其他传感器，它们都具有相同的基本结构：由目标化合物（直接或通过去除阻遏物）诱导的基因启动子，融合在报告基因的下游。后者可以编码可追踪的蛋白质（例如 GFP），或更常见的是编码酶，可以实时定量监测酶的活性（van der Meer 和 Belkin，2010）。必要时，还必须克隆调控元件，特别是当充当传感元件的基因启动子不是宿主生物体固有的时。鉴于实际上无限数量的基因启动子和调节蛋白可用作候选传感器元件，此类传感器可能的传感目标的范围异常广泛。在开发特定化合物的微生物传感器的同时，生物报告菌株也被描述用于检测全局样本特征，例如毒性或遗传毒性/致突变性、对环境健康以及化学品安全重要的参数。商业 SOS Chromotest（Quillardet 等人，1982）是这组测定的先驱，随后是 umu 测试（Oda 等人，1985）。在这两种情况下，DNA 损伤剂对大肠杆菌 SOS 修复调节子的基因启动子的激活均通过 lacZ 作为报告基因进行显色监测。回顾过去 15 年，微生物生物传感器设计最强大的创新可能是合成生物学时代的到来。虽然这个术语在一个多世纪前就被引入科学文献中（Leduc，1910），但多年来它的含义已经慢慢发生了变化。随着雅各布和莫诺模型的引入，微生物生物技术的视野随着越来越复杂的分子工具的出现而打开，其中包括来自不同微生物和病毒的大量酶，它们被利用和重新训练以执行剪切、粘贴和编辑技巧。当这些酶的嗜热变体被巧妙地用于 PCR 技术的发明时，同样的视野实际上爆炸了，当基因组测序变得琐碎和生物信息化时，同样的视野变得基本上无限......
It is tempting to speculate that sixty years ago, when Jacob and Monod presented their model of the lac operon (Jacob and Monod, 1961), they already had a glimpse of the future of the lacZ gene, not only as encoding a cleaver of disaccharides, nor as a component in a beautiful and groundbreaking model of gene regulation, but also as a universal reporter of gene activation. Indeed, reporter gene technology rapidly became a basic tool in studying the regulation of gene expression; several decades had to pass, however, before the same approach has led to the first report of a microorganism genetically engineered to perform an accurate, specific and sensitive analysis of an environmental pollutant (King et al., 1990). The term ‘whole cell biosensor’soon entered into use, accompanied by some semantic controversy: purists view the term ‘biosensor’as a hardware device, in which the biological entity (eg enzyme, antibody, oligonucleotide or a live cell) serves as its sensing component (IUPAC, 2017); according to this view, a microbial strain, notwithstanding the complexity of its re-engineering, may be called a ‘sensor strain’or a ‘bioreporter’, but never a ‘biosensor’. Long before this linguistic polemic became an issue, however, a pioneering article from the Sayler group (King et al., 1990) described a bioluminescent Pseudomonas-based sensor of naphthalene. This publication was trailed by the first E. coli-based mercury sensor (Selifonova et al., 1993), soon to be followed by numerous others, all sharing the same basic structure: a gene promoter induced by the target compound (directly, or via the removal of a repressor), fused downstream of a reporter gene. The latter could code for a traceable protein (eg GFP) or–more often–for an enzyme, the activity of which could be monitored quantitatively in real time (van der Meer and Belkin, 2010). When necessary, regulatory elements had to be cloned as well, especially when the gene promoter acting as the sensing element was not native to the host organism. In view of the practically infinite number of gene promoters and regulatory proteins available as candidate sensor elements, the scope of possible sensing targets of such sensors is exceptionally broad. In parallel to the development of microbial sensors of specific compounds, bioreporter strains have also been described for the detection of global sample characteristics such as toxicity or genotoxicity/mutagenicity, parameters of importance for environmental health as well as for chemicals’ safety. The commercial SOS Chromotest (Quillardet et al., 1982), the forerunner of this group of assays, was followed by the umu-test (Oda et al., 1985). In both cases, the activation of gene promoters from the E. coli SOS repair regulon by DNA damaging agents was chromogenically monitored with lacZ as a reporter gene.Looking back over the last 15 years, possibly the most powerful innovator of microbial biosensor design was the coming of age of synthetic biology. While the term has been introduced to the scientific literature over a century ago (Leduc, 1910), its meaning has slowly changed over the years. Following the introduction of the Jacob and Monod model, microbial biotechnology horizons opened up with the advent of increasingly more sophisticated molecular tools, including numerous enzymes derived from diverse microorganisms and viruses, harnessed and retrained to perform cutting, pasting and editing tricks. The same horizons practically exploded when thermophilic variants of these enzymes were ingeniously employed in the invention of PCR technology, and turned essentially limitless when genome sequencing was made trivial and bioinformatic …
登录
查看更多内容
影响因子：
64.8
作者：
Fredens, Julius;Wang, Kaihang;Chin, Jason W.
通讯作者：
Chin, Jason W.
影响因子：
17.1
作者：
Courbet, Alexis;Endy, Drew;Bonnet, Jerome
通讯作者：
Bonnet, Jerome
DOI：
10.1073/pnas.1918859117
发表时间：
2020-03-24
影响因子：
11.1
作者：
Fan, Catherine;Davison, Paul A.;Huang, Wei E.
通讯作者：
Huang, Wei E.
影响因子：
64.5
作者：
Pardee, Keith;Green, Alexander A.;Collins, James J.
通讯作者：
Collins, James J.
DOI：
10.1073/pnas.79.19.5971
发表时间：
1982-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
QUILLARDET, P;HUISMAN, O;HOFNUNG, M
通讯作者：
HOFNUNG, M