Kingston, Canada Queens_Canada iGEM 2011 Americas Jamboree content - PowerPoint PPT Presentation


 Kingston, Canada Queens_Canada iGEM 2011 Americas Jamboree content the problem The Challenge Our solution Approach & Methods Future Applications Accomplishments

one organism. one solution. degrade report toxins target chemical move to to Sustainable/ non-intrusive contaminated approach site NEMO REMEDIATION why C. elegans ? - Excellent(chemotaxis( mechanism( - Mul12cellular=(mul1ple( processes( - Hardy(organism(

part design Chemotaxis Parts *Teng et al., 2006; UCSF 2010 Promoter' Effect' Interchange(GPCR(genes( ODR7( A?rac1ve( STR2( A?rac1ve(( GYC7( Repulsive( GPC1( Repulsive( GPCR' Source' Ligand' ADBR2( Mouse( Phenol( Orexin( Human( Toluidine,(42Chlorobenzoic(acid,(( Naphthalene(( OX1R( Mouse( 42Chlorobenzoic(acid( MARM1( Rat( Phenol( NPYR1( Mouse( Naphthalene( chemotactic assay results - Conducted(preferen1al(assay(between(target(( Part: ODR-7:ADBR2:UNC-54 (BBa_K631032) and(control(substances( 1( 0.8( Chemotactic Index 0.6( 0.4( Transgenic Control 0.2( !"#$% 0( = # ! !"!#$ ! !"#$% ! !" ! !"#$%& ! !"#$%#&' ! − # ! !"!#$ ! !"#$% ! !" ! !"#$%"& ! !"#$%#&' ! 0.03( 0.05( 0.08( ! # ! !"!#$ ! !"#$% 20.2( Concentration (g/mL) !

naphthalene degradation naphthalene degradation Naphthalene(converted(to(salicylic(acid( - 22hydroxychromene222 carboxylate(isomerase(( - nahD( - Pseudomonas*pu,da ( part: BioBrick nahD future applications - Characterize(in( C.*elegans (( - Assembly(with(other(enzymes(in( C.*elegans ,( create(full(pathway( - BioBrick(Registry:(develop(bioremedia1on( pathways(for(other(chassis((( (

part improvement injection reporter - Modify(exis1ng(eCFP(part( - Test(new(promoters(effec1vely(( BioBrick Part: BBa_K6310
 C. elegans neurons GPC-1:eCFP:UNC-54:1C3 Expression in AWB neuron

kill switch mechanism conventional methods of BioBrick assembly standard assembly 3-way assembly Target(Gene( novel assembly: PCR ligation conventional method of BioBrick assembly 1C3 1A3 1T3

acknowledgements sponsors Faculty Advisors : Dr. Chin-Sang, Dr. Greer, Dr. Ko, Dr. Bendena, Dr. (( Ramsay, & Dr. Young. Lab Support : Tony Papanicolaou, Jeff Boudreau and rest of Chin-Sang lab. Past QGEM Team Members : Steve Goldie, Chris Palmer, Geoff Halliday, Harry Zhou and James MacLeod. QGEM 2011 : Adrian McNeely, Tony He, Stephanie Zhou, Olivia Margie, Alex Campbell, Allister Smith, David Dai, Sophie Gong, Kevin Chen, Kristina Collavino, Eni Rukaj, Faisal Bakhteyar, Eszter Gereb, Gary Wu, Wei Liu, Anujan Poologaindran. QGEM 2010/2011 Executive Team references 1.(Ahmed(S,(Hodgkin(J.((2000)(MRT22(checkpoint(protein(is(required(for(germline(immortality(and(telomere(replica1on(in( C.*elegans .(Nature,(403:1592164(( ( 2.(Smelick(C,(Ahmed(S.((2005)(Achieving(immortality(in(the( C.*elegans (germline.(Ageing(Research(Reviews,(4(1):67282(( ( 3.(Telomere(Shortening(and(Damage((2009)((h?p://www.immortalhumans.com/telomere2shortening2and2damage/)( ( 4.(Fire(A,(Xu(S,(Montgomery(MK,(Kostas(SA,(Driver(SE,(Mello(CC.((1998)(Potent(and(specific(gene1c(interference(by(double2stranded(RNA(in(Caenorhabdi1s( elegans.(Nature,(391(6669):806211( ( 5.(Tabara(H,(Grishok(A,(Mello(CC.((1998)(RNAi(in( C.*elegans :(soaking(in(the(genome(sequence.(Science,(282(5388):43021( ( 6.(Timmons(L,(Fire(A.((1998)(Specific(interference(by(ingested(dsRNA.(Nature,(395(6705):854( ( 7.(Timmons(L.((2006)(Construc1on(of(Plasmids(for(RNA(Interference(and(In(Vitro(Transcrip1on(of(Double2Stranded(RNA.(Methods(Mol(Biol.(351:109217.( ( 8.(Simmer(F,(Tijsterman(M,(Parrish(S,(Koushika(SP,(Nonet(ML,(Fire(A,(Ahringer(J,(Plasterk(RHA.((2002)(Loss(of(the(Puta1ve(RNA2Directed(RNA(Polymerase( RRF23(Makes(C.(elegans(Hypersensi1ve(to(RNAi.(Current(Biology,(12(15):(131721319.( ( 9.(Kiyohara(H,(Torigoe(S,(Kaida(N,(Asaki(T,(Iida,(T,(Hayashi(H(and(Takizawa(N.((1994)(Cloning(and(Characteriza1on(of(a(Chromosomal(Gene(Cluster,(pah,(That( Encodes(the(Upper(Pathway(for(Phenanthrene(and(Naphthalene(U1liza1on(by(Pseudomonas(pu1da(OUS82.(Journal(of(Bacteriology,(176:243922443.(( ( Graphics' (order(of(appearance)( h?p://s.ngm.com/2009/03/canadian2oil2sands/img/candian2oil2sands2615.jpg(( h?p://en.wikipedia.org/wiki/GPCR(( h?p://ginkgobioworks.com/support/BioBrick_Assembly_Manual.pdf( h?p://www.ncbi.nlm.nih.gov/pmc/ar1cles/PMC205370/((( h?p://www2fgg.eur.nl/ch1/cellbiology/images/jansen1.jpg((