SPS ingapore Controlled missiles: Targeted treatment of tumors with - PowerPoint PPT Presentation

SPS ingapore Controlled missiles: Targeted treatment of tumors with engineered E. coli

Team SPSingapore

In a Nutshell Biological Programming Simplifying Circuits In a Nutshell Biological Programming HP: Simplifying Circuits

In a Nutshell Biological Programming Simplifying Circuits

In a Nutshell Biological Programming Simplifying Circuits

In a Nutshell Biological Programming Simplifying Circuits

In a Nutshell Biological Programming Simplifying Circuits

In a Nutshell Biological Programming Simplifying Circuits Quorum Sensing O 2

In a Nutshell Biological Programming Simplifying Circuits Quorum Sensing Prevents bacterial activity unless sufficient bacteria are present

In a Nutshell Biological Programming Simplifying Circuits O 2 O 2 Anaerobic switch O 2 Prevents bacterial activity outside of the hypoxic tumour region O 2 O 2 O 2

In a Nutshell Biological Programming Simplifying Circuits O 2 O 2 O 2 QS OFF QS ON Anaerobic ON Anaerobic OFF No expression No expression

In a Nutshell Biological Programming Simplifying Circuits 13 Quorum Anaerobic sensing switch

In a Nutshell Biological Programming Simplifying Circuits In a Nutshell Biological Programming HP: Simplifying Circuits

In a Nutshell Biological Programming Simplifying Circuits 15 Quorum Anaerobic sensing switch

In a Nutshell Biological Programming Simplifying Circuits 16 Quorum Anaerobic sensing switch aaaaaaaaaa

In a Nutshell Biological Programming Simplifying Circuits nirB /FNR anaerobic system • nirB promoter is anaerobic-responsive

In a Nutshell Biological Programming Simplifying Circuits 18 Quorum Anaerobic sensing switch

In a Nutshell Biological Programming Simplifying Circuits EsaR/EsaI quorum sensing system

In a Nutshell Biological Programming Simplifying Circuits 20 Quorum Anaerobic sensing switch

In a Nutshell Biological Programming Simplifying Circuits Invasin + Listeriolysin effector proteins • Updated documentation of BBa_K299812 • Sequence differs significantly from parts registry Invasin • BioBricks suffix is missing in construct (Pstl only) Listeriolysin

In a Nutshell Biological Programming Simplifying Circuits Invasin + Listeriolysin effector proteins • Updated documentation of BBa_K299812 • Listeriolysin is ‘codon - optimised’ • Invasin is 300bp longer • Invasin matches Yersinia enterocolitica Documented Invasin Listeriolysin 4146bp in parts registry Sequenced Invasin +300bp Listeriolysin 4460bp from sequencing

In a Nutshell Biological Programming Simplifying Circuits Invasin + Listeriolysin • Constitutive expression of Invasin and Listerolysin (From BBa_K299812) Mammalian cell P lac Listeriolysin Invasin Evaluate ability of part to P nirB Invasin confer invasion phenotype and escape from endosome Positively-regulated by FNR protein in Anaerobic conditions

In a Nutshell Biological Programming Simplifying Circuits Testing our tools: P nirB- Invasin vector • There was increased invasion under anaerobic conditions

In a Nutshell Biological Programming Simplifying Circuits Testing our tools: P lac -GFP • P lac promoter shows high basal expression Green fluorescence GFP P lac detectable at 488nm

In a Nutshell Biological Programming Simplifying Circuits Testing our tools: P lac -GFP Untransformed E. coli pSB1C3-P lac GFP

In a Nutshell Biological Programming Simplifying Circuits Submissions to parts registry Registry ID Name of Construct Purpose BBa_K299812 pSB1A2-inv-hly Sequence confirmed sample BBa_K1804001 pSB1C3-p lac- gfp Constitutively expressing GFP BBa_K1804002 pSB1C3-p nirB P nirB Anaerobic promoter

In a Nutshell Biological Programming Simplifying Circuits In a Nutshell Biological Programming HP: Simplifying Circuits

In a Nutshell Biological Programming Simplifying Circuits 30 Explaining our system Quorum Anaerobic sensing switch

In a Nutshell Biological Programming Simplifying Circuits Genetic Circuits • Informative, but unappealing

In a Nutshell Biological Programming Simplifying Circuits Brute-force representation

In a Nutshell Biological Programming Simplifying Circuits Simplified genetic circuits • A teaching tool to introduce genetic circuit designing • Simplified abstraction of genetic circuit elements

In a Nutshell Biological Programming Simplifying Circuits

In a Nutshell Biological Programming Simplifying Circuits Workshop use • To test the tool, we integrated it into a workshop. • Presented it as a set of puzzles • Varying levels of difficulty • Multiple ways of obtaining the solution

In a Nutshell Biological Programming Simplifying Circuits Workshop • One-day workshop • Mostly first-year biology students, and some participants from other disciplines • Theoretical and experimental components

In a Nutshell Biological Programming Simplifying Circuits Introduction & Theory Quick introduction to genetics Genetic circuit puzzles

In a Nutshell Biological Programming Simplifying Circuits Wet lab: Fusion PCR Basic Laboratory Training Introduction to gel electrophoresis

In a Nutshell Biological Programming Simplifying Circuits Wet lab: Bacteria with GFP Viewing GFP-transformed bacteria Microscope view

In a Nutshell Biological Programming Simplifying Circuits After the workshop • Most enjoyed the workshop, with puzzles being the most popular. • Non-biology majors picked up on the puzzles fairly quickly. • Participants displayed good conceptual understanding.

In a Nutshell Biological Programming Simplifying Circuits After the workshop • We also sought further feedback from educators. • Received interest in developing it for science outreach • Ideally we would like to develop the tool further, and make it open-access.

In a Nutshell Biological Programming Simplifying Circuits What we’ve done this summer • Designed a dual-switch system • Submitted three parts to the BioBricks registry • Developed an educational tool on genetic circuits • Conducted a workshop testing the tool

THE END We would like to thank: Special Programme in Science Faculty of Science, National University of Singapore Mentors: Dr. Leslie Gapter, Prof. Linda Kenney, Dr. Stuti Desai Integrated DNA Technologies