P RIS Bettencourt TUBERCULOSIS IS A WORLDWIDE DISEASE 1 3 OF - - PowerPoint PPT Presentation

Bettencourt

P RIS

TUBERCULOSIS IS A WORLDWIDE DISEASE

OF THE WORLD POPULATION HAS LATENT TUBERCULOSIS

1 3

MILLION

1.3 DEATHS PER YEAR

detect infjltrate target sabotage gender study

FIGHT TUBERCULOSIS MODERN WEAPONS

with

INFILTRATE

detect infjltrate target sabotage gender study

IT MOSTLY AFFECTS THE LUNGS ...

LATENT TB FORMS GRANULOMAS IN THE LUNGS WHERE IT CAN SURVIVE EVEN INSIDE MACROPHAGES

TUBERCULOSIS CAN BE

ACTIVE LATENT

OR

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

TDMH-expressing E.coli kill mycobacteria in culture 2 4 6 10

5

10

6

10

7

Mycobaterial Survivors Time (Hours)

Mycobacteria alone Mycobacteria +

• E. coli (Uninduced)

Mycobacteria +

• E. coli (TDMH-induced)

BIOBRICKED

C O L L A B O R A T I O N

X 1

DETECT

detect infjltrate target sabotage gender study

TO DIAGNOSE MULTI DRUG RESISTANCE CAN TAKE FROM

> 21 DAYS TO 3 MONTHS <

gRNA

reporter

reporter Cas9

Antibiotic Resistance

LexA

!

SOS response causes

Xgal

Blue!

lacZ pRecA

Lex A

Chromosome

DETECT

A biosensor that detects the presence of specifjc antibio- tic resistance genes

DETECT

10

2

10

3

10

4

10

5

10

6

Surviving transformants (CFU/ml) C A S 9 g R N A B

• t

h

KanR- KanR+

Cotransformation of E. coli with Cas9 and gRNA vectors in Kanamycin Sensitive and Resistant Strains BIOBRICKED

ASSEMBLY STANDARD

Parallel assembly of genes of interest TOOLS

{

{

BioBrick Backbone Multiple inserts

SP SP S P EX E X EX Complete BioBrick Plasmid

SENSIGEM : A BIOSENSOR DATABASE

www.sensigem.org

C O L L A B O R A T I O N

iGEM

2013

OPENDATA TOOLS

SENSIGEM : A BIOSENSOR DATABASE

(6

Nucleic Acid Biosensors (6) Other Biosensors (221)

Underrepresentation of nucleic acid biosensors

GENDER STUDY

detect infjltrate target sabotage gender study

35% OF ADULTS INFECTED WITH TB ARE WOMEN

BIOLOGICAL SOCIAL

vt

Our Advisors & Instructors Our Team

? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?

BIOLOGICAL SOCIAL

vt

Our Team Our Advisors & Instructors

? ? ? ?

BIOLOGICAL SOCIAL

vt

Our Advisors & Instructors Our Team

DATA DRIVEN APPROACH

1Database 50 ALL

731

Synthetic Biology Laboratories SB Conferences Teams screened All info available on wiki

OPENDATA

WOMEN ARE UNDER REPRESENTED SYNTHETIC BIOLOGY AND

in

2007 2009 2011

0%

20% 40% 60% 80% 100%

Women Men

The iGEM gender gap has not changed signifjcantly since 2007 Gender Balance of Synthetic Biology Labs

60 40 20 Labs Phd students Post Doc Head of Labs

Percent of iGEMers Year

What did you hope to learn in iGEM

WOMEN DO NOT SUPERVISE MEN

as much as

0% 10% 20% 30% 40% 50% Percent Women Students Supervisors Judges *

Sex ratio in iGEM according to role

1 2 3 4 5 Working on a team Designing a project General lab experience Human practices Programming and Modeling Leading a team Leading a project

Men Women

Interest level (5 = high)

DIVERSITY SUCESS

Women are better represented in winning iGEM teams

0% 10% 20% 30% 40% 50%

*

p = 0.035

Percent women Prize - winners All teams

iGEM a leader and an example of an active gender policy that can be measured

RECOMMENDATIONS

Raise the number of women judges Motivate young women to be advisors Encourage gender refmection in SynBio

SABOTAGE

detect infjltrate target sabotage gender study

MDR PATIENT ARE

SUCCESSFULLY

TREATED

1 3

SABOTAGE

CmR

Add sRNA

P h a g e

LB+Cm LB

Plate:

Antibiotic!

A phage system with low fjtness cost producing sRNA which inhibit the synthesis of antibiotic resistance proteins

SABOTAGE

Killing Chloramphenicol resistant E.Coli population with Chloramphenicol Origin of resistance of the sequential killing

Concentration of Chloramphenicol (ug/mL) % of surving cells Non infected Infected

Non silenced Non infected

50 500 1000

SABOTAGE

Adaptable silencing tool

Pr promoter Scaffold Originated from MicC T1/TE 24 bp Target-binding sequence

Anti-Cm

E P S X

BIOBRICKED

TARGET

detect infjltrate target sabotage gender study

ONE NEW DRUG APPROVED IN THE LAST 40 YEARS

TARGET

WT mycoSIR E.coli

Compare Growth Inhibition

FIND NEW DRUG!

A safe and specifjc high throughput drug screening method that targets essential mycobacteria metabolic proteins

TARGET

Growth curves of E. coli mycoSIR BIOBRICKED

3 6 9 12 0.1 0.2 0.3 0.4

A

Time (hours) Growth (OD)

0.1 0.2 0.3 0.4

B

WT BL21 MycoSIR E.coli MycoSIR uninduced

BL21 cysl

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

detect infjltrate target sabotage

SCIENTIFIC ACCOMPLISHMENTS

detect infjltrate target sabotage

4 new biobricks New assembly standard Detection of kanamycin resistance gene

SCIENTIFIC ACCOMPLISHMENTS

detect infjltrate target sabotage

4 new biobricks New assembly standard Detection of kanamycin resistance gene 1 new biobrick E.coli that kills

• M. smegmatis

SCIENTIFIC ACCOMPLISHMENTS

detect infjltrate target sabotage

4 new biobricks New assembly standard Detection of kanamycin resistance gene 1 new biobrick E.coli that kills

• M. smegmatis

3 new biobricks Successful silenced antibiotic resistance cassettes Modelling

4 new biobricks New assembly standard Detection of kanamycin resistance gene 3 new biobricks Functional metabolic pathway of M. smegmatis in E. coli Modelling 3 new biobricks Successful silenced antibiotic resistance cassettes Modelling 1 new biobrick E.coli that kills

• M. smegmatis

SCIENTIFIC ACCOMPLISHMENTS

detect infjltrate target 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 75014 Paris FRANCE +33 1 44 41 25 22/25 team2013@igem-paris.org

Bettencourt

P RIS

THE TEAM

A Lindner J Wintermute M Toulouze Z Marinkovic

Our Advisors & Instructors Our Team

I Aghoghogbe

C Basier

E Atanasković

M Deyell

M Benony

N Koutsoubelis

S Zahra

S Jaramillo R. V Libis

A Bernheim

Y Zegman

A Löchner C Bencherif

detect infjltrate target sabotage gender study

FIGHT TUBERCULOSIS MODERN WEAPONS

with