Institut Clinique de la Souris - NIH Common Fund · Institut Clinique de la Souris. Contribution to the IMPC . ... HEMMERLE Mathieu JACQUOT Sylvie ... PHAM Thi Bich Hanh POUILLY Laurent

Institut Clinique de la Souris

The French Contribution to the IMPC

Yann Herault CNRS, INSERM, Université de Strasbourg,

1 rue Laurent Fries - 67404 Illkirch, France. [email protected]

The French National Infrastructure for Mouse PHENOGENOMICS

M. Selloum, M.-C. Birling C. Olivier, S. Lerondel F. Fiore, B. Malissen, G. Pavlovic, A. Ayadi, P. Lopes, C. Fremond, T. Sorg, Y. Herault

TAAM

ICS CIPHE

WP1: Management

USE

RS/

PAR

TNER

S

WP2

Mutagenesis WP3

Zootechny WP4

Phenotyping

USE

RS/

PAR

TNER

S

WP5: Discover of mammalian genes function /

Contribution to IMPC

1. ES Cells to Mouse 180 40 30

2. Mouse to Cohort 180 40 30

3. Cohort to Phenotype 250 250

4. Archive and Distribute 180 40 30

5. Tech and Tools Dev 1,2,3 4 1,3

promoter

start codon

stop codon

prom-Neo

FRT loxP FRT loxP loxP SA-lacZ-pA

Cre recombinase cre

promoter

start codon

stop codon

loxP FRT SA-lacZ-pA

lacZ

Critical exon

tm1a

tm1b (post-Cre)

KO first allele (reporter-tagged insertion allele)

QC and procedures from ES to cohorts

•• southsouthern ern blot blot anaanalylysis sis ffor ror rececombinaombination tion ccononfirmafirmation tion as as wwellell asasccononfirmafirmation tion of tof the he prpresence esence of the of the 3’ 3’ llooxPxP sitsite,e,

•• kkararyyotyotyping of 2 vping of 2 validaalidatted ed clonesclones•• micrmicroo--injection injection in 50 in 50 BBALB/ALB/cNcN blasblasttocyocytteses•• esesttablishablishmemennt t of of chimerchimeraass•• brbreeding eeding of the of the 4 bes4 best t chimerchimeras as ((chimerichimerismsm >45%)>45%)•• GLGLT assessT assessmemennt t (tm1a(tm1a allelallelees s ffor or crcryyoo))•• GenerGeneraation tion of tm1b of tm1b KKO allele O allele (pos(postt--CrCree))•• IVF IVF quick quick eexpansionxpansion HeHet t (4 males and 8 females)•• HeHet t x Hex Het t crcrossoss HomHom (4 males and 4 females)•• HomHom X HeX Het t crcrosses: osses: prproducoduction oftion of ccohoohort, frt, fertiliertility ty and and viviabiliabilityty

ttesests ts (n=4)(n=4 Males and females)•• 1 Cohort 1 Cohort of cof conontrtrol per wol per weekeek

Genetic Tools Dev.: Speeding up the Cre-Step with Fluo marker

a b

c d

Rosa26Cre-GFP/0

Rosa26Flp-YPF/0

B6N

M-C. Birling, G. Pavlovic

Cre eGFP Neo 2A

FlpO eYFP pA Neo 2A pCAGGS

pChick b-act

Rosa26 WT

Rosa26Cre-GFP(FRT-Neo-FRT)

Rosa26FLP-YFP(LoxP-Neo-LoxP)

pA

PCR1

PCR4 PCR2

Cre eGFP 2A pChick b-act pA

PCR4 PCR3

Rosa26Cre-GFP(FRT)

PCR5 PCR7

FlpO eYFP pA

CR7

2A

P

Maternal Rosa26 allele

WT Flp-YFP(loxP-Neo-loxP) P

ater

nal

Ro

sa2

6

alle

les

WT

Cre-GFP (FRT-Neo-FRT)

WT/WT

14

Maternal contribution (FRT)/Cre-GFP WT

10

WT/Flp-YFP(LoxP-Neo-loxP)

13

Cre-GFP (FRT) /Flp-YFP(loxP)

7

Maternal Rosa26 allele

WT Cre-GFP (FRT-Neo-FRT)

Pate

rnal

Ro

sa2

6

alle

les WT

WT/WT

29

WT/Cre-GFP(FRT-Neo-FRT)

28

Flp-YFP (LoxP-Neo-loxP)

Maternal contribution

Flp-YFP (loxP)/WT

28

Flp-YFP(loxP)/Cre-GFP (FRT)

13

pCAGGS

Rosa26FLP-YFP(LoxP)

PCR6

Genetic Tools Dev.: and Maternal Cre effect Rosa26Cre-GFP(FRT)-/Flp-YFP(LoxP)

1 2 3 4 5 6 7

Rosa26Cre-GFP(FRT-Neo-FRT)/ WT

1 2 3 4 PCR

LoxP FRT

M-C. Birling, G. Pavlovic

16

9 13 10 11 14 15 12

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TECH DEVELOPMENT Cardiovascular

Behavior and cognition Nutrition, metabolism

Neuromuscular system

Embryology and Reproduction

Immunology and Inflammation

Respiration, allergy Imaging immune responses to pathogens

Skeleton, Tooth and Craniofacial morphology

Cytometry by time-of-flight mass spectrometry Electrophysiology

Scientific Experts: Block-Zupan A., Chambon P., Chan S., Fiore F., Frossard N., Gorvel J.-P., Grenot P., Hérault Y., Kieffer B., Laporte J., Lepape A., Malissen B., Malissen M., Mark M., Metzger D., Monassier L.,Ouagazzal A.M., Pourquié O., Ricci R., Romand R., Roux M., Spitz F., Ugolini S., Vivier E., …

Follow gestation by micro-ultrasonography

at E5 and E8 (count the deciduas)

No or few (less than 40 %) resorptions

Collect fetuses at E18.5

Externally malformed fetuses

Fix 3 litters in Bouin’s solution

No mutant detected

Collect embryos at E10.5

Externally normal fetuses

Fix 3 litters in Klotz’s solution

No mutant detected

Collect embryos at E10.5

Genotyping

Mutants detected

Autopsy (Klotz) or Serial sections (Bouin)

High level of resorptions

Collect embryos at E10.5

Fix 2 litters in paraformaldehyde

b-galactosidase expression by optical

tomography

Genotyping

No mutant detected

End of analysis (Death must occur around implantation or gastrulation)

Fix 2 litters in Bouin’s fluid

Mutants detected

Serial sections

Gestation at E7

44/105

O. Wendling, M. Mark

TECH DEVELOPMENT Cardiovascular

Behavior and cognition Nutrition, metabolism

Neuromuscular system

Embryology and Reproduction

Immunology and Inflammation

Respiration, allergy Imaging immune responses to pathogens

Skeleton, Tooth and Craniofacial morphology

Cytometry by time-of-flight mass spectrometry Electrophysiology

Scientific Experts: Block-Zupan A., Chambon P., Chan S., Fiore F., Frossard N., Gorvel J.-P., Grenot P., Hérault Y., Kieffer B., Laporte J., Malissen B., Malissen M., Mark M., Metzger D., Monassier L., Ouagazzal A.M., Pourquié O., Ricci R., Romand R., Roux M., Spitz F., Ugolini S., Vivier E., …

Landmark and methodologies for craniofacial analysis

L. Viriot, P. Lopes, E. Velot, S. Lerondel, Y. Herault

TECH DEVELOPMENT Cardiovascular

Behavior and cognition Nutrition, metabolism

Neuromuscular system

Embryology and Reproduction

Immunology and Inflammation

Respiration, allergy Imaging immune responses to pathogens

Skeleton, Tooth and Craniofacial morphology

Cytometry by time-of-flight mass spectrometry Electrophysiology

Scientific Experts: Block-Zupan A., Chambon P., Chan S., Fiore F., Frossard N., Gorvel J.-P., Grenot P., Hérault Y., Kieffer B., Laporte J., Lepape A., Malissen B., Malissen M., Mark M., Metzger D., Monassier L., Ouagazzal A.M., Pourquié O., Ricci R., Romand R., Roux M., Spitz F., Ugolini S., Vivier E., …

Lipopolysaccharide (LPS) challenge

0,00

2,00

4,00

6,00

8,00

10,00

0 100 200 300 400 500

ctrl

LPS 1µg

LPS 5µg

LPS 10 µg

BALF Serum BALF Serum

Pulmonary and inflammatory Function (6h) E. Dalloneau, J. Becker, Y. Herault

Lipopolysaccharide (LPS) challenge

Prmt2 NF-kB

TLR4

TNFα IL-6

LPS

0

1

2

3

4

5

6

52

03

55

06

58

09

51

10

12

51

40

15

51

70

18

52

00

21

52

30

24

52

60

27

52

90

30

53

20

33

5 min

Pulmonary airway resistance WT NaCl

Prmt2+/- NaCl

WT LPS

Prmt2+/- LPS

0

200

400

600

800

1000

pg/

ml

[TNF] **

**

NaCl LPS

E. Dalloneau, J. Becker, Y. Herault (J. Immunol in press)

0

400

800

1200

1600

2000

pg

/ml

[IL-6] **

**

NaCl LPS

TECH DEVELOPMENT Cardiovascular

Behavior and cognition Nutrition, metabolism

Neuromuscular system

Embryology and Reproduction

Immunology and Inflammation

Respiration, allergy Imaging immune responses to pathogens

Skeleton, Tooth and Craniofacial morphology

Cytometry by time-of-flight mass spectrometry Electrophysiology

Scientific Experts: Block-Zupan A., Chambon P., Chan S., Fiore F., Frossard N., Gorvel J.-P., Grenot P., Hérault Y., Kieffer B., Laporte J., Lepape A., Malissen B., Malissen M., Mark M., Metzger D., Monassier L., Ouagazzal A.M., Pourquié O., Ricci R., Romand R., Roux M., Spitz F., Ugolini S., Vivier E., …

Profiling of the mouse innate and adaptive immune system under normal and confined

(BSL3) conditions via the quantitative analysis of over 200 parameters.

Mass Cytometer

INSERM US12

Standard operating procedure for quantitative analysis of the lymphoid and myeloid cells

present in small blood samples.

Standard operating procedure for

quantitative analysis of dendritic cells.

REACHING THE COMMUNITY > 1000 Lab Users

CIPHE ICS

TAAM

European funded networks

21

1953-2011: 58 years serving the scientific community

BERTEAUX LIONEL

BERTRAND MARIE-FRANCOISE

BILAN Christiane

BORNAT ANGELIQUE

BOULANGER VIRGINIE

CHAVANEAU GHISLAINE

CHERFOUH AMEZIANE

CLEMENCON RUDY

DESALE ELODIE

DESSAIN MARIE LAURE

DHENNIN LAURENT

DHENNIN RAPHAELLE

DIET Alexandre

DOUIN Estelle

DUFOUR ALAIN

DURAND TAMARA

FOURNIER KARINE

FREMOND CECILE

FROUX NATHALIE

HERPIN Alexandre

JAMBOU KARINE

JOSEPH Laurent

JOUSSARD GUILLAUME

LANDREAU CATHERINE

LARRIGALDIE VANESSA

LE BRIS KHEIRA

LE MEE MARYLINE

LERONDEL STEPHANIE

LORY RACHEL

MAILLARD Karine

MAISTRE Jean-baptiste

MARTIN Christelle

MAUNY PASCAL

MAURIO AGNES

MEAUXSOONE JOEL

MEDINA PATRICE

MERCIER LUDOVIC

MOTHU Nadya

MOUSSU JEAN-PAUL

PAPION DUCHATEAU RENE

PATURANCE SEBASTIEN

PERES PHILIPPE

RENIER ANNICK

RETIF LEROUX STEPHANIE

SEGHROUCHNI EMMANUELLE

SENECHAL ARMELLE

THERET YVETTE

THOMAS Jean-Luc

TORSET MONIQUE

VOISIN OLIVIER

ZIADI ABDELMALEK

LERONDEL STEPHANIE

LORY RACHEL

MAILLARD Karine

http://transgenose.cnrs-orleans.fr/taam/presentation.php

Please visit our new website http://www.ics-mci.fr/

ALI-HADJI Dalila

AMANN Grégory

ANDRE Philippe

AUBURTIN Aurélie

AUGE Fabrice

AYADI Abdelkader

BAM'HAMED Chaouki

BANQUART- OTT Nadine

BECKER Julien

BEDU Elodie

BIRLING Marie-Christine

BLONDELLE Eric

BOUR Raphaël

BRIGNON Sophie

BUNZ Isabel

CARADEC Claudia

CAYROU Pauline

CES Aurélia

CHAMPY Marie-France

CHARLES Philippe

CHARTOIRE Nathalie

CHEBBOUB Djaouida

CLAEYSSEN Richard

COMBE Roy

DEBOUZY Guillaume

DELANGLE Benoît

DIERICH Andrée

DREYER Dominique

EGLINGER Yolande

EL FERTAK Leila

EL FERTAK Lahcen

ENNAH Hamid

ERBS Valérie

ESSABRI Karim

FISCHER Fabienne

FISCHER Natacha

FOUGEROLLE Jean-Victor

FRICKER Bastien

GOETZ-REINER Patrice

GONCALVES DA CRUZ Isabelle

GRUBER Frédéric

GUIMOND Alain

HERAULT Yann

HEMMERLE Mathieu

JACQUOT Sylvie

JAGLA-EBERLIN Monika

KUJATH Christelle

KURTZ Caroline

LAEUFER Laurent

LALANNE Valérie

LE MARCHAND Elise

LEBLANC Sophie

LECOCQ Muriel

LEGEAY Sandrine

LEYRITZ Johan

LINDNER Loic

LORENTZ Romain

LUPPI Laurence

LUX Aline

MELLUL Peggy

MERTZ Annelyse

MEZIANE Hamid

MITTELHAEUSER Christophe

MOKNI Mourad

MONTIAL Marina

MORO Anne-Isabelle

MOULAERT David

MULLER Stéphanie

NAGRE Isabelle

OHLMANN Thierry

PAVLOVIC Guillaume

PENSAVALLE Joëlle

PERRY Mélanie

PETER Emilie

PETIT-DEMOULIERE Benoît

PHAM Thi Bich Hanh

POUILLY Laurent

QUEUCHE Danielle

RIET Fabrice

RIO-ZENNER Fanny

ROTH Christelle

ROUSSEAU Stéphane

ROUSSEAU Valérie

SCHMITT Raphaël

SCHOEDEL Christophe

SCHWOERER Marie-Jeanne

SEITZ Thierry

SELLOUM Mohammed

SORG-GUSS Tania

SUTTER WOLTER Anne

TILLY Isabelle

TOUBARI Chadia

UZUN Ibrahim

VASSEUR Laurent

VENTEO Lydie

VINCENT Cindy

WAGNER Christel

WALCH Laëtitia

WALLERICH Sandrine

WEBER Bruno

WENDLING Olivia

WETZSTEIN Eric

WIECROCK Cyrille

WIECROCK Ludovic

ZANINELLO Fabienne

ZANINELLO Nathalie