Biofilm Lithography enables high-resolution cell patterning via optogenetic adhesin expression.
Biofilm Lithography enables high-resolution cell patterning via optogenetic adhesin expression.
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DOI：
10.1073/pnas.1720676115
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发表时间：
2018-04-03
影响因子：
11.1
通讯作者：
Riedel-Kruse IH
中科院分区：
文献类型：
--
作者：
Jin X;Riedel-Kruse IH
细菌生活在被称为生物膜的表面附着的群落中，那里的空间结构与群落功能紧密相连。我们已经开发了一种通过工程细菌进行基因编码的生物膜图案化工具(“生物膜光刻”)，从而使负责表面附着的膜黏附蛋白的表达受到光学调控。因此，这些细菌只在光照的表面区域形成生物膜。利用这一工具，我们能够使用蓝光以25μm的空间分辨率对大肠杆菌生物膜进行图案绘制。我们提出了一个附带的生物物理模型，以了解光调节生物膜形成背后的机制，并对相关的自然生物膜过程提供洞察。总体而言，这种生物膜图案化工具可以用于研究自然微生物群落，也可以用于设计活的生物材料。细菌生物膜为微生物群落工程提供了一个很有希望的机会。然而，我们控制生物膜空间结构的能力仍然有限。在这里，我们利用光激活转录启动子(PDawn)来光学调节粘附素基因(Ag43)的表达。在有图案的蓝光照射下，可以在不同的基质和封闭的培养室内形成空间分辨率低至25μm的长期存活的生物膜，而不需要表面处理。生物物理模型表明，图案化机制涉及刺激瞬时表面吸附的细胞，为先前提出的粘附素在自然生物膜成熟过程中的作用提供了证据。总体而言，这种被称为“生物膜光刻”的工具比现有的细胞沉积/图案化方法具有明显的优势，提供了生长结构化生物膜的能力，应用于更好地了解自然生物膜群落，以及生物材料工程和自下而上的微生物联合体设计方法。
Bacteria live in surface-attached communities known as biofilms, where spatial structure is tightly linked to community function. We have developed a genetically encoded biofilm patterning tool (“Biofilm Lithography”) by engineering bacteria such that the expression of membrane adhesion proteins responsible for surface attachment is optically regulated. Accordingly, these bacteria only form biofilm on illuminated surface regions. With this tool, we are able to use blue light to pattern Escherichia coli biofilms with 25 μm spatial resolution. We present an accompanying biophysical model to understand the mechanism behind light-regulated biofilm formation and to provide insight on related natural biofilm processes. Overall, this biofilm patterning tool can be applied to study natural microbial communities as well as to engineer living biomaterials. Bacterial biofilms represent a promising opportunity for engineering of microbial communities. However, our ability to control spatial structure in biofilms remains limited. Here we engineer Escherichia coli with a light-activated transcriptional promoter (pDawn) to optically regulate expression of an adhesin gene (Ag43). When illuminated with patterned blue light, long-term viable biofilms with spatial resolution down to 25 μm can be formed on a variety of substrates and inside enclosed culture chambers without the need for surface pretreatment. A biophysical model suggests that the patterning mechanism involves stimulation of transiently surface-adsorbed cells, lending evidence to a previously proposed role of adhesin expression during natural biofilm maturation. Overall, this tool—termed “Biofilm Lithography”—has distinct advantages over existing cell-depositing/patterning methods and provides the ability to grow structured biofilms, with applications toward an improved understanding of natural biofilm communities, as well as the engineering of living biomaterials and bottom–up approaches to microbial consortia design.
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影响因子：
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