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research article

Engineering Agrobacterium tumefaciens Adhesion to Target Cells

Pierrat, Xavier  
•
Pham, Alix  
•
Wong, Jeremy P. H.
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July 26, 2022
Acs Synthetic Biology

Agrobacterium tumefaciens is a plant pathogen commonly repurposed for genetic modification of crops. Despite its versatility, it remains inefficient at transferring DNA to many hosts, including to animal cells. Like many pathogens, physical contact between A. tumefaciens and host cells promotes infection efficacy. Thus, improving the strength and specificity of A. tumefaciens to target cells has the potential for enhancing DNA transfer for biotechnological and therapeutic purposes. Here, we demonstrate a methodology for engineering genetically encoded exogeneous adhesins at the surface of A. tumefaciens. We identified an autotransporter gene we named Aat that is predicted to show canonical beta-barrel and passenger domains. We engineered the beta-barrel scaffold and linker (Aat(beta)) to display synthetic adhesins susceptible to rewire A. tumefaciens to alternative host targets. As a proof of concept, we leveraged the versatility of a VHH domain to rewire A. tumefaciens adhesion to yeast and mammalian hosts displaying a GFP target receptor. Finally, to demonstrate how synthetic A. tumefaciens adhesion can improve transfer to host cells, we showed improved protein translocation into HeLa cells using a sensitive split luciferase reporter system. Engineering A. tumefaciens adhesion has therefore a strong potential in generating complex heterogeneous cellular assemblies and in improving DNA transfer efficiency against non-natural hosts.

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Type
research article
DOI
10.1021/acssynbio.2c00069
Web of Science ID

WOS:000834177500001

Author(s)
Pierrat, Xavier  
Pham, Alix  
Wong, Jeremy P. H.
Al-Mayyah, Zainebe  
Persat, Alexandre  
Date Issued

2022-07-26

Publisher

AMER CHEMICAL SOC

Published in
Acs Synthetic Biology
Subjects

Biochemical Research Methods

•

Biochemistry & Molecular Biology

•

t-dna transfer

•

surface display

•

outer-membrane

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protein

•

secretion

•

vire2

•

coli

•

transformation

•

efficiency

•

selection

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
UPPERSAT  
Available on Infoscience
August 15, 2022
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/189963
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