A riboswitch controlled location- based networking framework - PowerPoint PPT Presentation

A riboswitch controlled location- based networking framework

Bacterial chemotaxis

Chemotaxis: chez-mediated flagellar rotation

o vervi erview: ew: de design ign Simulation ation Ch Characteri acterizatio zation Outr treach ach Summar mary y & ac achievements vements Acknowle owledge dgeme ments nts TH AN K YO U

END CheZ-reporter START Design Construction Function

Project Design & Construction Analyte A Analyte A Analyte B gradient gradient START Analyte B

towards A Analyte A CheZ-Venus Send & Receive motility module P on CheZ-mRFP CheZ-Venus START lox66 lox71 Analyte B Riboswitch A Riboswitch B A B

towards A Analyte A CheZ-Venus P on CheZ-mRFP lox66 CheZ-Venus START lox71 Analyte B Riboswitch A Riboswitch B Cre-activating A B toggle switch Y P X X Y Cre P Y X

towards B Analyte A CheZ-mRFP P on lox71 CheZ-mRFP lox66 CheZ-Venus START Analyte B Riboswitch A Riboswitch B B A Cre Y P X X Y Cre P Y X

Atra razine zine rib iboswitch switch Atraz azin ine e ribos oswitc witch (Sinha ha and and Gallivan livan, NCB NCB , , 2010) 10)

Riboswitches Theophyll hylline ne ribosw switch itch 1 8.1 Theophy phylli line ne Ribos oswitch witch (Topp pp and and Galliva ivan, , JACS , , 2007) )

Theophyll hylline ne ribosw switch itch 2 12.1 1 Theophy phylline lline ribos oswitc witch (Lynch ch and Gallivan ivan, , NAR, 2009)

11nt insert theophylline riboswitch BBa_K249028 Lethbridge 6nt scar theophylline riboswitch BBa_K411001 Taipei Venus mRFP CheZ CDS “x” WITS_CSIR thRS-CheZ-reporter

Overvie rview s im imulation ulation Ch Characteri acterizatio zation Outr treach ach Summar mary y & ac achievements vements Acknowle owledge dgeme ments nts TH AN K YO U

Overvie rview: w: design gn Simulation ation c ha haract racterizat erization ion Outr treach ach Summar mary y & ac achievements vements Acknowle owledge dgeme ments nts TH AN K YO U

Empty vector Riboswitch 1 Riboswitch 2 RS1 Venus RS2 Venus BBa_K537009 BBa_K537010

8-fold activation Riboswitch 1 chez venus RS1 CheZ Venus BBa_K537011

46-fold activation RS2 CheZ Venus Riboswitch 2 chez venus K537012

Riboswitch 1 CheZ BBa_K537011 CheZ RS1 Venus Riboswitch 2 CheZ BBa_K537012 RS2 CheZ Venus

BBa_K537012 5-fold difference BBa_K537011

Riboswitch 1 Riboswitch 2

Overview rview Simulation ation Ch Characteri acterizatio zation Outreach utreach Summar mary y & ac achievements vements Acknowle owledge dgeme ments nts Th ank Yo u

Wor orksho kshops ps at Sci-bon bono o disc scove very y centre ntre: “national science week” 2-6 6 Au Augus ust 2011 11

Problem Instruction + ‘Superhero’ bacteria! Solution

Overview rview Simulation ation Ch Characteri acterizatio zation Outr treach ach S ummar ary y & a & achie ievem vemen ents Acknowle owledge dgeme ments nts TH AN K YO U

Introduced synthetic biology to the broader public Submitted 11 new parts to the registry Fully characterized two theophylline riboswitches Collaborated with ICL on modelling and assays

A finalist at the European Jamboree Improved the aesthetics and content of wiki Improved experiments & microscopy Had lots of fun!

Overview rview Si Simulation lation Ch Characteri acterizatio zation Outr treach ach Summar mary y & ac achievements vements Ackn knowledgement owledgements THANK YOU

• Prof Kramer Supervisors • Prof Patrick Arbuthnot • Karl Rumbold • Prof Theresa Coetzer • Marco Weinberg • Dr Sylvia Fanucchi • Musa Mhlanga • Prof Heini Dirr • Raymond Sparrow • David Balchin • Robyn Brackin • Advisors The Synthetic Biology Research Unit at the CSIR • Michelle Robinson • Dr Sonja Lauterbach • Laura Millroy • Kubendran Naidoo • Youtaro Shibayama • Dr Toru Nakayashiki and Prof Hirotada Mori • Dr Joy Sinha and Prof Justin Gallivan