Kingston, Canada Queens_Canada iGEM 2011 Americas Jamboree content - - PowerPoint PPT Presentation
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
NEMOREMEDIATION move to to contaminated site report toxins degrade target chemical Sustainable/ non-intrusive approach
- ne organism. one solution.
why C. elegans?
- Excellent(chemotaxis(
mechanism(
- Mul12cellular=(mul1ple(
processes(
- Hardy(organism(
part design
Interchange(GPCR(genes(
*Teng et al., 2006; UCSF 2010
Chemotaxis Parts
Promoter' Effect' 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(
- Conducted(preferen1al(assay(between(target((
and(control(substances(
chemotactic assay
!"#$% = #!!"!#$!!"#$%!!"!!"#$%&!!"#$%#&'! − #!!"!#$!!"#$%!!"!!"#$%"&!!"#$%#&'! #!!"!#$!!"#$% ! !
results
Part: ODR-7:ADBR2:UNC-54 (BBa_K631032)
20.2( 0( 0.2( 0.4( 0.6( 0.8( 1(
0.03( 0.05( 0.08( Chemotactic Index Concentration (g/mL)
Transgenic Control
naphthalene degradation
Naphthalene(converted(to(salicylic(acid(
naphthalene degradation
- 22hydroxychromene222
carboxylate(isomerase((
- nahD(
- Pseudomonas*pu,da(
part: BioBrick nahD
- Characterize(in(C.*elegans((
- Assembly(with(other(enzymes(in(C.*elegans,(
create(full(pathway(
- BioBrick(Registry:(develop(bioremedia1on(
pathways(for(other(chassis((( (
future applications
part improvement
- Modify(exis1ng(eCFP(part(
- Test(new(promoters(effec1vely((
injection reporter
- C. elegans neurons
BioBrick Part: BBa_K6310 GPC-1:eCFP:UNC-54:1C3 Expression in AWB neuron
kill switch mechanism
Target(Gene(
conventional methods of BioBrick assembly
standard assembly 3-way assembly
conventional method of BioBrick assembly
1C3 1T3 1A3
novel assembly: PCR ligation
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
acknowledgements
((
sponsors
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((