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Title:A genetically encoded system for programmable cell assembly using surface-displayed DNA oligonucleotides
Author(s):Min, Yuhao
Contributor(s):Zhao, Huimin
Subject(s):ssDNA
cell surface
genetic
assembly
Abstract:Precise patterning of cells in three dimensions is highly desirable in both basic and applied research for tissue engineering. Although bottom-up cell assembly of with defined spatial arrangements has been achieved using cell-surface-bound DNA oligonucleotides, the current approaches require complicated chemical functionalization protocols, which is expensive, time-consuming and difficult to scale up. To overcome these limitations, we devised a genetically encoded system to attach customized single-stranded DNA (ssDNA) sequences onto Escherichia coli cell surface. In particular, in vivo produced ssDNA molecules containing a specific recognition site could be covalently bound to a phage protein, X174 A*, which was fused with a membrane protein as the cell surface anchor. As proof-of-concept, recombinant expression cassettes of X174 A* and the ssDNA biosynthesis machinery were constructed and introduced in E. coli, and the presence of surface-attached ssDNA molecules was examined using flow cytometry. Employing a cell-impermeable ssDNA-binding fluorescence dye, we observed fluorescence signals only when both X174 A* and ssDNA-synthesizing proteins were expressed. In addition, fluorescence was diminished after treatment using a ssDNA-specific exonuclease, confirming the successful display of ssDNAs on the cell surface. The feasibility of using this system for programmable assembly of E. coli cells is under investigation.
Issue Date:2016
Citation Info:Min, Yuhao. (2016, April). "A genetically encoded system for programmable cell assembly using surface-displayed DNA oligonucleotides." Session presented at Undergraduate Research Symposium, University of Illinois at Urbana-Champaign, Illinois.
Genre:Conference Poster
Type:Text
Language:English
URI:http://hdl.handle.net/2142/90235
Sponsor:IGB
Rights Information:Copyright 2016 Yuhao Min
Date Available in IDEALS:2016-06-07


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