1. Biofilm Lithography enables high-resolution cell patterning via optogenetic adhesin expression
- Author
-
Xiaofan Jin and Ingmar H. Riedel-Kruse
- Subjects
0301 basic medicine ,Light ,030106 microbiology ,High resolution ,Optogenetics ,medicine.disease_cause ,Microbiology ,biofilm ,Bacterial Adhesion ,03 medical and health sciences ,Escherichia coli ,medicine ,Lithography ,Adhesins, Escherichia coli ,Multidisciplinary ,bacterial patterning ,Spatial structure ,Chemistry ,pDawn-Ag43 ,Biofilm ,Biological Sciences ,biochemical phenomena, metabolism, and nutrition ,Cell patterning ,Cell biology ,Bacterial adhesin ,030104 developmental biology ,Biofilms ,lithography - Abstract
Significance 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.
- Published
- 2018