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Barcoding biological reactions with DNA-functionalized vesicles

Justin A. Peruzzi, Miranda L. Jacobs, Timothy Q. Vu, View ORCID ProfileNeha P. Kamat
doi: https://doi.org/10.1101/672287
Justin A. Peruzzi
aDepartment of Chemical and Biological Engineering Northwestern University
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Miranda L. Jacobs
bDepartment of Biomedical Engineering, Northwestern University, McCormick School of Engineering, Technological Institute, 2145 Sheridan Road, Evanston Il, 60208
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Timothy Q. Vu
bDepartment of Biomedical Engineering, Northwestern University, McCormick School of Engineering, Technological Institute, 2145 Sheridan Road, Evanston Il, 60208
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Neha P. Kamat
bDepartment of Biomedical Engineering, Northwestern University, McCormick School of Engineering, Technological Institute, 2145 Sheridan Road, Evanston Il, 60208
cCenter for Synthetic Biology, Northwestern University
dChemistry of Life Processes Institute, Northwestern University
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  • ORCID record for Neha P. Kamat
  • For correspondence: nkamat@northwestern.edu
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Abstract

Targeted vesicle fusion is a promising approach to selectively control interactions between vesicle compartments and would enable the initiation of biological reactions in complex aqueous environments. Here, we explore how two features of vesicle membranes, DNA tethers and phase-segregated membranes, promote fusion between specific vesicle populations. We show that membrane phase-segregation provides an energetic driver for membrane fusion that increases the efficiency of DNA-mediated fusion events. Using this system, we show that orthogonality provided by DNA tethers allows us to direct fusion and delivery of DNA cargo to specific vesicle populations. We then demonstrate that vesicle fusion between DNA-tethered vesicles can be used to initiate in vitro protein expression that leads to the synthesis of model soluble and membrane proteins. The ability to engineer orthogonal fusion events between DNA-tethered vesicles will provide a new strategy to control the spatio-temporal dynamics of cell-free reactions, expanding opportunities to engineer artificial cellular systems.

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Posted June 15, 2019.
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Barcoding biological reactions with DNA-functionalized vesicles
Justin A. Peruzzi, Miranda L. Jacobs, Timothy Q. Vu, Neha P. Kamat
bioRxiv 672287; doi: https://doi.org/10.1101/672287
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Barcoding biological reactions with DNA-functionalized vesicles
Justin A. Peruzzi, Miranda L. Jacobs, Timothy Q. Vu, Neha P. Kamat
bioRxiv 672287; doi: https://doi.org/10.1101/672287

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