mouse_tail, Illustration of mouse's tail that transforms into a strand of DNA, Credit to Darryl Leja, NHGRI December 3, 2002 

Mouse Genetics and Inflammation

Laboratory

 
 

UniTag

30/05/2018


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Practical Course SS 2018 04.06.-20.07.2018

Masters Degree (M.Sc.), Module MN-B-SM (G1): Mouse Genetics, Cell Death and Inflammation

Introduction to the module:


30th of  May 2018, 13:00, Institut für Genetik, seminar room 0.46, ground floor. 

Course overview:

During this course you will participate in two practical parts (I +II), a theoretical week with Journal Club, Lectures and a final exam.

04/06/- 15/06/2018 Practical part I:


“Engineering the mouse genome with CRISPR/Cas and site specific recombinases”

18/06/- 22/06/2018 Journal Club Week:

You will get a brief introduction to  NF- k B signalling and you will be given original research papers describing key findings on the function and regulation of the  NF- k B pathway. You will spend this week with careful critical reading of the research papers, literature searches to obtain background information and in depth discussion of the manuscripts with assigned tutors, who will advise you on how to assemble a 'journal club' presentation. At the end of that week all participants in the course will present their papers in a 'journal club', which is part of the course evaluation process.

11.06.18 Introduction to NF- k B signalling, 09:00,  Institut für Genetik , seminar room 0.46, ground floor.

22.06.18 Jounral Club from 9:00 on,  CECAD, ground floor,  EG.037/038 .

25/06/- 13/07/2018 Practical part II:

"Mouse Genetics as a tool to investigate signalling cascades in vivo"
 
The practical parts consits of lectures and practical training in the laboratory. If not anounced differently the lectures start 9:00, Institut für Genetik, seminar room 0.46, ground floor. (EXEPTION: ON FRIDAYS and ON TUESDAY 5th of June the lectures take place in seminar rom 4th floor!!!) )

Lectures:

  1. Mon, 04.06.18 CRISPR/Cas (Laurens) followed by the safety instruction (Martin Hafner)
  2. Tue, 05.06.18 Modern Mouse Genetics I ( Prof. Manolis Pasparakis, 9:00 seminar room 4th floor, Inst. for Genetics)
  3. Wed, 06.06.18 Nuclease assisted Gene Tergeting (Laurens)

 

 

  1. Mon, 11.06.18 Introduction to NF- k B signalling (Prof. Manolis Pasparakis)
  2. Tue, 12.06.18 NF- k B mediated transcriptional regulation (Teresa)  

 

Cell Death Pathways

  1. Wed, 13.06.18 Caspases (Prof. Hamid Kashkar)
  2. Thr,14.06.18 Apoptosis, External and Internal Pathways (Prof. Hamid Kashkar)
  3. Fri, 15.06.18 TNF signalling (Prof. Manolis Pasparakis, 9:00 seminar room 4th floor, Inst. for Genetics)
  4. Mon,18.06.18 Necroptosis ( Prof. Manolis Pasparakis)

 

Immunity and Inflammation

  1. Mon, 25.06.18 Introduction: Immune System ( Prof. Manolis Pasparakis)
  2. Tue, 26.06.18 Innate Immunity ( Prof. Manolis Pasparakis)
  3. Wed, 27.06.18 TLR signalling (Prof. Manolis Pasparakis)  
  4. Thr, 28.06.18 Inflammasomes ( Prof. Hamid Kashkar )

 

NF- k B in inflammatory diseases: Lessons from mouse model

  1.   Mon, 02.07.18 Inflammatory Bowel Disease (Robin)
  2.   Tue, 03.07.18 Inflammatory Liver Disease (t.b.a)
  3.   Wed, 04.07.18 Inflammatory Skin Disease (Sneha)
  4.   Thr, 05.07.18 Cell Death in Bacterial Infection (Prof. Hamid Kashkar)

 

  1. Mon, 09 .07.18 Immunohistochemistry
     
    Friday, 13.07.18 reports results (from 9:00 on), CECAD, ground floor,  EG.037/038 .


    Exam: 20.07.18, 10:00-12:00, Institut für Genetik , lecture hall ground floor.

    Protocol due: 17.08.18

    Practical Course instructors:

    • Teresa Corona, Claudia Uthoff-Hachenberg Laurens Wachmuth together with other members of the Pasparakis Lab
    • ...

    Reading List:

    Capecchi M.R. Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century. Nat Rev Genet. 2005 Jun;6(6):507-12)

    Lewandoski M. Conditional control of gene expression in the mouse. (Nat Rev Genet. 2001 Oct;2(10):743-55.)

    Oeckinghaus A., Hayden M.S. and Ghosh S. Crosstalk in NF-κB signaling pathways. (Nat Immunol. 2011 Jul 19;12(8):695-708.)

    The new frontier of genome engineering with CRISPR-Cas9 GENOME EDITING. J.A. Doudna and E. Charpentier. (Science. 2014 Nov 28;346(6213))

    Necroptosis and its role in inflammation. Pasparakis M., Vandenabeele P. (Nature. 2015 Jan 15;517(7534):311-20. Review.)

    Literature for the theoretical part:  

    RIPK1 counteracts ZBP1-mediated necroptosis to inhibit inflammation.
    Lin J, Kumari S, Kim C, Van TM, Wachsmuth L, Polykratis A, Pasparakis M. Nature. 2016 Dec 1;540(7631):124-128.

    NEMO Prevents Steatohepatitis and Hepatocellular Carcinoma by Inhibiting RIPK1 Kinase Activity-Mediated Hepatocyte Apoptosis.   Kondylis V, Polykratis A, Ehlken H, Ochoa-Callejero L, Straub BK, Krishna-Subramanian S, Van TM, Curth HM, Heise N, Weih F, Klein U, Schirmacher P, Kelliher M, Pasparakis M., Cancer Cell. 2015 Nov 9;28(5):582-98.

    Phosphorylation-Driven Assembly of the RIP1-RIP3 Complex Regulates Programmed Necrosis and Virus-Induced Inflammation . YoungSik Cho, Sreerupa Challa, David Moquin, Ryan Genga, Tathagat Dutta Ray, Melissa Guildford, and Francis Ka-Ming Chan.   Cell 137, 1112–1123, June 12, 2009 ª.

    LUBAC-Recruited CYLD and A20 Regulate Gene Activation and Cell Death by Exerting Opposing Effects on Linear Ubiquitin in Signaling Complexes. Draber P, Kupka S, Reichert M, Draberova H, Lafont E, de Miguel D, Spilgies L, Surinova S, Taraborrelli L, Hartwig T et al. Cell Reports 13, 2258–2272, December 15, 2015.

    FADD prevents RIP3-mediated epithelial cell necrosis and chronic intestinal inflammation. Welz PS, Wullaert A, Vlantis K, Kondylis V, Fernández-Majada V, Ermolaeva M, Kirsch P, Sterner-Kock A, van Loo G, Pasparakis M. Nature. 2011 Jul 31;477(7364):330-4.

    IAP antagonists target cIAP1 to induce TNFalpha-dependent apoptosis. Vince JE1, Wong WW, Khan N, Feltham R, Chau D, Ahmed AU, Benetatos CA, Chunduru SK, Condon SM, McKinlay M, Brink R, Leverkus M, Tergaonkar V, Schneider P, Callus BA, Koentgen F, Vaux DL, Silke J. Cell. 2007 Nov 16;131(4):682-93. .

    BID-dependent release of mitochondrial SMAC dampens XIAP-mediated immunity against Shigella. Andree M1, Seeger JM1, Schüll S1, Coutelle O1, Wagner-Stippich D1, Wiegmann K2, Wunderlich CM3, Brinkmann K1, Broxtermann P1, Witt A1, Fritsch M1, Martinelli P4, Bielig H2, Lamkemeyer T5, Rugarli EI4, Kaufmann T6, Sterner-Kock A7, Wunderlich FT3, Villunger A8, Martins LM9, Krönke M1, Kufer TA2, Utermöhlen O10, Kashkar H11. EMBO J. 2014 Oct 1;33(19):2171-87. .

    Links concerning the practical course:

    Results Practical:

    Western p-IkBa:

    • Group1
    • Group2
    • Group3
    • Group4
    • Group5
    • Group6
    • Group7
    • Group8

     

    GFP p65 MEFs

    • group1
    • group2
    • group3
    • group4
    • group5
    • group6
    • group7
    • group8

     

    FACS:

    • group1
    • group2
    • group3
    • group4
    • group5
    • group6
    • group7
    • group8

     

    PCRs

    • group1
    • group2
    • group3
    • group4
    • group5
    • group6
    • group7
    • group8

     

    Longterm stimulation microscopy

    • groups 1,3,5,7
    • groups 2,4,6,8

    H & E

    • group1
    • group2
    • group3
    • group4
    • group5
    • group6
    • group7
    • group8

     

    CD3 stainings

    • group1
    • group2
    • group3
    • group4
    • group5
    • group6
    • group7
    • group8

    Please check again for more information!

    Last update 30/05/18