A genetically encoded system for programmable cell assembly using surface-displayed DNA oligonucleotides

Min, Yuhao

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https://hdl.handle.net/2142/90235 Copy

Description

Title

A genetically encoded system for programmable cell assembly using surface-displayed DNA oligonucleotides

Author(s)

Min, Yuhao

Contributor(s)

Huimin, Zhao

Issue Date

2016

Keyword(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.

Type of Resource

text

Language

en

Permalink

http://hdl.handle.net/2142/90235

Sponsor(s)/Grant Number(s)

IGB

Copyright and License Information

Copyright 2016 Yuhao Min

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