P RIS Bettencourt
TUBERCULOSIS IS A WORLDWIDE DISEASE 1 3 OF THE WORLD POPULATION HAS LATENT TUBERCULOSIS MILLION 1.3 DEATHS PER YEAR
FIGHT TUBERCULOSIS MODERN WEAPONS with target detect infjltrate sabotage gender study
INFILTRATE target detect IT MOSTLY AFFECTS THE LUNGS ... TUBERCULOSIS CAN BE OR ACTIVE LATENT LATENT TB FORMS GRANULOMAS IN THE LUNGS WHERE IT CAN SURVIVE EVEN INSIDE MACROPHAGES infjltrate sabotage gender study
INFILTRATE M. smegmatis Cytoplasm TDMH LLO LLO : Listeriolysin O TDMH : Trehalose Dimycolate Hydrolase
INFILTRATE Infjltrating macrophages with an E.coli producing TDMH - an E.coli that lyses mycobacterial cell walls
INFILTRATE BIOBRICKED 1 X TDMH-expressing E.coli kill mycobacteria in culture N O I T A R O B A L L O C Mycobaterial Survivor s Mycobacteria 7 10 alone Mycobacteria + E. coli (Uninduced) 6 10 Mycobacteria + E. coli (TDMH-induced ) 5 10 0 2 4 6 Time (Hours)
DETECT target detect TO DIAGNOSE MULTI DRUG RESISTANCE CAN TAKE FROM > 21 DAYS TO 3 MONTHS < infjltrate sabotage gender study
DETECT A biosensor that detects the presence of specifjc antibio - tic resistance genes Antibiotic Resistance reporter gRNA Chromosome Cas9 SOS response causes LexA cleavage LexA ! reporter Lex A Blue! pRecA lacZ Xgal
DETECT BIOBRICKED X4 Cotransformation of E. coli with Cas9 and gRNA vectors in Kanamycin Sensitive and Resistant Strains 6 10 Surviving transformants (CFU/ml) 5 10 4 10 3 10 KanR- 2 KanR+ 10 A h 9 t N o S R B A g C
ASSEMBLY STANDARD TOOLS Parallel assembly of genes of interest BioBrick Backbone { SP EX Multiple inserts { EX Complete BioBrick SP Plasmid S X P E
SENSIGEM : A BIOSENSOR DATABASE N www.sensigem.org O I T A R O B A L L O C iGEM 2013 OPENDATA TOOLS
SENSIGEM : A BIOSENSOR DATABASE Underrepresentation of nucleic acid biosensors (6 Nucleic Acid Biosensors (6) Other Biosensors (221)
GENDER STUDY target detect 35% OF ADULTS INFECTED WITH TB ARE WOMEN infjltrate sabotage gender study
BIOLOGICAL SOCIAL vt Our Team ? ? ? ? ? ? ? ? ? ? ? ? ? Our Advisors & Instructors ? ? ? ?
BIOLOGICAL SOCIAL vt Our Team Our Advisors & Instructors ? ? ? ?
BIOLOGICAL SOCIAL vt Our Team Our Advisors & Instructors
DATA DRIVEN APPROACH OPENDATA 50 Synthetic Biology 1 Database Laboratories ALL SB Conferences 731 All info available on wiki Teams screened
WOMEN ARE UNDER REPRESENTED SYNTHETIC BIOLOGY AND in Gender Balance of The iGEM gender gap has not changed Synthetic Biology Labs signifjcantly since 2007 100% 60 Men 80% Percent of iGEMers 40 60% 40% 20 Wome n 20% 0 0% 2007 2009 2011 Labs Phd Post Head of student s Doc Labs Year
WOMEN DO NOT SUPERVISE MEN as much as Sex ratio in iGEM according to role What did you hope to learn in iGEM Men 5 Women 50% * Interest level (5 = high) 4 40% Percent Women 30% 3 20% 2 10% 0% 1 Working on a team Designing a projec t General lab experienc e Programming and Modeling Human practices Leading a team Leading a projec t Student s Supervisors Judges
DIVERSITY SUCESS Women are better represented in winning iGEM teams * 50% 40% Percent women 30% 20% 10% p = 0.035 0% Prize - All teams winners
RECOMMENDATIONS Raise the number of women judges Motivate young women to be advisors Encourage gender refmection in SynBio iGEM a leader and an example of an active gender policy that can be measured
SABOTAGE target detect 1 MDR PATIENT ARE SUCCESSFULLY 3 TREATED infjltrate sabotage gender study
SABOTAGE A phage system with low fjtness cost producing sRNA which inhibit the synthesis of antibiotic resistance proteins Add sRNA P Plate: h a g e LB M 1 3 P M13 Phage h a g e Antibiotic! CmR A N R s LB+Cm Antibiotic resistance Shhh!!!
SABOTAGE Killing Chloramphenicol resistant E.Coli Origin of resistance of the sequential population with Chloramphenicol killing % of surving cells Non infected Non silenced 0 50 500 1000 Concentration of Chloramphenicol (ug/mL) Non infected Infected
SABOTAGE BIOBRICKED X3 Adaptable silencing tool 24 bp Target-binding sequence S P E X Scaffold Originated T1/TE P r promoter Anti-Cm from MicC
TARGET target detect ONE NEW DRUG APPROVED IN THE LAST 40 YEARS infjltrate sabotage gender study
TARGET A safe and specifjc high throughput drug screening method that targets essential mycobacteria metabolic proteins WT mycoSIR E.coli SirA FprA FdxA FprA 3NADPH3 NADP FdxA FdxA RED OX CysI CysI SirA NO Growth High High Throughout Throughout Screening Screening Compare Growth Inhibition FIND NEW DRUG!
TARGET BIOBRICKED Growth curves of E. coli mycoSIR X3 A B 0.4 0.4 0.3 0.3 Growth (OD) 0.2 0.2 0.1 0.1 0 0 BL21 cysl MycoSIR WT BL21 MycoSIR E.coli uninduced 0 3 6 9 12 Time (hours)
TECHNOLOGY TRANSFER Science and technology always have a human face! Focus on the concrete effects and local circumstances Talk to patients, doctors and policy makers Nicola Bertoldi
SCIENTIFIC ACCOMPLISHMENTS target detect infjltrate sabotage
SCIENTIFIC ACCOMPLISHMENTS 4 new biobricks New assembly standard Detection of kanamycin target detect resistance gene infjltrate sabotage
SCIENTIFIC ACCOMPLISHMENTS 4 new biobricks New assembly standard Detection of kanamycin target detect resistance gene 1 new biobrick E.coli that kills M. smegmatis infjltrate sabotage
SCIENTIFIC ACCOMPLISHMENTS 4 new biobricks New assembly standard Detection of kanamycin target detect resistance gene 3 new biobricks Successful silenced 1 new biobrick antibiotic resistance cassettes E.coli that kills M. smegmatis Modelling infjltrate sabotage
SCIENTIFIC ACCOMPLISHMENTS 3 new biobricks 4 new biobricks Functional metabolic New assembly standard pathway of M. smegmatis Detection of kanamycin target in E. coli detect resistance gene Modelling 3 new biobricks Successful silenced 1 new biobrick antibiotic resistance cassettes E.coli that kills M. smegmatis Modelling infjltrate sabotage
Centre for Research and Interdisciplinarity (CRI) Faculty of Medicine Cochin Port-Royal, South wing, 2nd fmoor Paris Descartes University 24, rue du Faubourg Saint Jacques P RIS 75014 Paris Bettencourt FRANCE +33 1 44 41 25 22/25 team2013@igem-paris.org
THE TEAM Our Team S Jaramillo R. Y Zegman N Koutsoubelis A Bernheim M Benony S Zahra C Basier M Deyell A Löchner C Bencherif V Libis I Aghoghogbe E Atanasković Our Advisors & Instructors A Lindner J Wintermute Z Marinkovic M Toulouze
FIGHT TUBERCULOSIS MODERN WEAPONS with target detect infjltrate sabotage gender study
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