Cell Surface Targeting (we

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Cell Surface Targeting(were actually trying it!)

• Lewis Hahn
• Perry Tsai
• June 26, 2006

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Outline

• Expressing Streptavidin at the E. coli cell
  surface
• Two display options
• Why it might work! (More about streptavidin)
• Creating a DNA-DNA interface with the cell
  surface
• What others have done
• Our plans

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Expressing Streptavidin at the E. coli cell
surface
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Two display options

• GPI-anchor
• Beta autotransporter

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GPI-anchor

• Jung et al. 1998
• expressed levansucrase (400 amino acids)
• Fused to C-terminus of Inp

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Autodisplay

• Jose 1997 (and 1999,2000,2001,)
• Beta autotransporter
• Originally used with AIDA-I (adhesin-involved-in-d
  iffuse-adherence)

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Autodisplay mechanism
(Jose 1996)
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Why it might work
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Creating a DNA-DNA interface with the cell
surface
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Previous work

• Tahiri-Alaoui et al. 2002

RNA-RNA
Adaptamer
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CopA-CopT
(Kolb 2000)
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CopA-CopT
(Kolb 2000)
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Many options

• To start work in solution.
• New interfaces?
• Secondary structure design?
• New Targets?

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Other targets

• Dwarakanath et al. 2002
• DNA Aptamer to LPS

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Summary

• A lot of building blocks are already there
• Streptavidin fusion protein
• GPI anchor
• Beta-autotransporter
• DNA-DNA or RNA-RNA interface
• Adaptamer
• New interfaces, new aptamers

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Major Question

• Will people talk to us?