Location & dates EMBL Heidelberg, Germany 16 - 21 Sep 2018
Deadlines Application closed
Organiser_ThermoFisher

Programme

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Sunday 16 September 2018
TimeSpeaker
from 17.00 Arrival & Welcome at the ISG Hotel

Welcoming round of organizers, trainers, speakers at the ISG Hotel

Ice-breaker (Flash talks)

18:30 - 19:15

Introduction into CRISPR/Cas9
Sibylle Chantal Vonesch - EMBL Heidelberg, Germany

19:30

Dinner - ISG Hotel

Monday 17 September 2018
TimeSpeaker
9.00 - 9.30

Arrival & Welcome
Yvonne Yeboah, Elisabeth Zielonka - EMBL Heidelberg, Germany

9.30 - 10.15 Purification of Cas9 protein
Kim Remans - EMBL Heidelberg, Germany
10.15 - 10.45 Coffee Break
10.45 - 11.45

gRNA/Donor design (Intro to practical)
Thermo Fischer Scientific

11.45 - 12.30 Practical 1-1
Guide RNA Synthesis – PCR for IVT templates
12.30 - 13.30 Lunch in the EMBL Canteen
13.30 - 14.30

CRISPR issues and applications
Dana Carroll - The University of Utah, United States of America

14.30 - 15.00

Coffee break

15.00 - 16.30

Practical 1-2
PCR gel analysis and set up IVT

16.30 - 18.00

Practicals 2 & 3 in groups
Transfection of 293FT cells with gRNA & Cas9mRNA lipofection
Electroporation of 293FT with gRNA & Cas9protein
Guide RNA design task

18.00 - 19.00

Dinner in the EMBL Canteen

19.00 - 20.30

Practicals 2 & 3 in groups continues
Transfection of 293FT cells with gRNA & Cas9mRNA lipofection
Electroporation of 293FT with gRNA & Cas9protein
Guide RNA design task

Tuesday 18 September 2018
TimeSpeaker
9.00 - 9.45

Genome and epigenome editing using CRISPR-Cas system
Claudio Mussolino - Medical Center -University of Freiburg, Germany

9.45 - 10.00

Coffee break

10.00 - 11.00 Generation and validation of endogenously tagged cell lines
Birgit Koch - Max Planck Institute for Medical Research, Germany
11.00 - 12.30 Practicals in groups
a) Introduction to FACS by Malte Paulsen
    FACS sorting of knock in cells – single cell sort
b) Individual project discussion
12.30 - 13.30 Lunch in the EMBL Canteen
13.30 - 15.00 Practicals in groups continues
a) Introduction to FACS by Malte Paulsen
    FACS sorting of knock in cells – single cell sort
b) Individual project discussion
15.00 - 15.15 Coffee break
15.15 - 17.45

Practicals 1-3 & 4 in groups continues
IVT cleanup, quantitation and analysis of gRNA
Transfection of 293FT-eBFP cells using CRISPR/Cas9 for SNP (RNPs)

17.45 - 18.30

Dinner in the EMBL Canteen

18.30 - 20.30

Practicals 1-3 & 4 in groups
IVT cleanup, quantitation and analysis of gRNA
Transfection of 293FT-eBFP cells using CRISPR/Cas9 for SNP (RNPs)

Wednesday 19 September 2018
TimeSpeaker
09.00 - 09.30

CRISPR & HiBiT Protein Tag –A perfect team to investigate endogenous protein biology
Erik Bonke - Promega, Germany

09.30 - 10.00 High Efficiency Gene Editing using Programmable EnGen Ribonucleoprotein Complexes
Karl von Laer - New England Biolabs, Germany
10.00 - 10.15

Coffee break

10.15 - 12.00

Practical 5-1 & 5-2
GCD Cell Harvest and lysis, set up GCD PCR

12.00 - 13.00

Lunch in the EMBL Canteen

13.00 - 13.45 Mammalian cell engineering: The genome editing toolbox and considerations for use
Jon Chesnut - Thermo Fisher Scientific, USA
13.45 - 14.15 Practical 5-3
CR analysis, set up GCD re-annealing reaction
14.15 - 16.15 Practicals 5-4 & 5-5 in groups
Set up digestion for GCD, gel purification & sample submission for sanger sequencing
Gel analysis of GCD
16.15 - 16.30 Coffee break
16.30 - 18.30

TIDE - rapid, powerful and easy analysis of CRISPR experiments
Victor Dillard - Desktop Genetics, UK

18.30 - 19.30 Dinner in the EMBL Canteen
Thursday 20 September 2017
TimeSpeaker
9.00 - 10.30 Practicals 6 in groups
Examine BFP cells via microscopy and FACS
Knock-In cell validation - DNA Preparation
10.30 - 12.00 Practicals 6 in groups continues
Examine BFP cells via microscopy and FACS
Knock-In cell validation - DNA Preparation
12.00 - 13.00 Lunch in the EMBL Canteen
13.00 - 13.45

Genome-wide and targeted screens using CRISPR/Cas9 system
Balca R. Mardin - BioMed X, Germany

13.45 - 15.00 Practical
Knock-In cell validation – Set up Junction PCR
15.00 - 15.30 Coffee Break
15.30 - 16.30 Optimizing CRISPR ribonucleoprotein components for precision genome editing
Justin Barr - Integrated DNA Technologies, United States of America
17.00 Shuttle downtown
from 19.00 Dinner downtown
Friday 21 September 2018
TimeSpeaker
9.00 - 10.00 CRISPR-Cas9 transgenic mice - from concept to founder
Neil Humphreys – EMBL Rome
10.00 - 11.00 Practical
Knock-In cell validation (1) - Gel electrophoresis of junction PCR
11.00 - 11.30

Coffee Break

11.30 - 12.15 Practical
Knock-In cell validation (2) - Analysis of gel electrophoresis
12.15 - 13.00 Final Project Discussion
13.00 - 14.00 Lunch in the EMBL Canteen
14.00 - 16.00

Summary of results, final discussions, feedback session

16.00 - 16.15

Coffee Break

16.15 - 16.30 Closing remarks & End of the course