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Cell Surface Targeting (we’re actually trying it!)

Cell Surface Targeting (we’re actually trying it!). Lewis Hahn Perry Tsai June 26, 2006. 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

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Cell Surface Targeting (we’re actually trying it!)

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  1. Cell Surface Targeting(we’re actually trying it!) Lewis Hahn Perry Tsai June 26, 2006

  2. 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

  3. Expressing Streptavidin at the E. coli cell surface

  4. Two display options • GPI-anchor • Beta autotransporter

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

  6. Autodisplay • Jose 1997 (and 1999,2000,2001,…) • Beta autotransporter • Originally used with AIDA-I (adhesin-involved-in-diffuse-adherence)

  7. Autodisplay mechanism (Jose 1996)

  8. Why it might work

  9. Creating a DNA-DNA interface with the cell surface

  10. Previous work • Tahiri-Alaoui et al. 2002 RNA-RNA “Adaptamer”

  11. CopA-CopT (Kolb 2000)

  12. CopA-CopT (Kolb 2000)

  13. Many options • To start: work in solution. • New interfaces? • Secondary structure design? • New Targets?

  14. Other targets • Dwarakanath et al. 2002 • DNA Aptamer to LPS

  15. 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

  16. Major Question Will people talk to us?

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