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Exploring Cell Surface Targeting: Streptavidin Display on E. coli

This presentation discusses the innovative approach of expressing streptavidin on the surface of E. coli, focusing on two display options: GPI-anchoring and beta-autotransporters. We explore why this method might succeed, the creation of DNA-DNA interfaces, and insights from previous studies in RNA-RNA systems. With a host of potential building blocks available, including streptavidin fusion proteins, we examine possibilities for advancing research on new aptamers and targets. Our key question: Will our findings encourage collaborative dialogue?

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Exploring Cell Surface Targeting: Streptavidin Display on E. coli

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Presentation Transcript

  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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