CB 399: Gene Targeting Strategies and Techniques Spring 2011

Intellectual Unit :

Gene Targeting Strategies and Techniques
Course Lecturers: Silvia Calpe, Research Fellow at the Division of Immunology at HMS and To-Ha Thai, Instructor, Harvard Medical School

Curriculum Fellows: Catherine Dubreuil, catherine_dubreuil@hms.harvard.edu and Latishya Steele, latishya_steele@hms.harvard.edu

This nanocourse will focus on the different strategies to generate genetically engineered mice. We will start describing the characteristics and methods of culturing embryonic stem cells and the strategies to generate gene targeted mice: knockout and knock-in mice. We will then discuss the generation and application of transgenic animals and its main applications. We will also discuss how both gene targeting and transgenesis techniques can be combined using the Cre-LoxP system in the generation of complex animal models.

Finally, we will introduce you to all available online resources and provide a strategic framework to follow in order to design your next “genetically engineered mouse”. In the second session we will discuss the applications of each of these technologies and how these technologies can be implemented in your research.


First Session: Wednesday, April 13, 2011, 2-5 pm
Location: TMEC Building, Walter Amphitheater

Second Session: Wednesday, April 20, 2011, 2-5 pm
Location: TMEC Building, Rm. 333


Assignment for the second session

Please choose one of the following options:

In a 5-minute in-class presentation (chalk talk or PowerPoint format, your choice):

Option 1: Select one article from the groups below (to download the .pdfs, you must be logged into nanos and quarters). Explain why the technique(s) were used in the studies presented in the article you chose. In your presentation, be sure to also mention if there is a better method the authors could have used and why you think this might be the case.

Option 2: Apply the material from the first session to your own research: if you had to create a murine model to address a question in your project, which method(s) might you use and why? If you choose this method, please send a short description of your research project to Sylvia Calpe (scalpe_f@bidmc.harvard.edu) by 5 pm on Monday, April 18.



Papers for Option 1 :

Group 1: Cell Ablation and Cell Activation

  • Haak et al. IL-17A and IL-17F do not contribute vitally to autoimmune neuro-inflammation in mice. J Clin Invest. 2009 Jan;119(1):61-9. doi: 10.1172/JCI35997. Epub 2008 Dec 15. 

Group 2: Conditional and Cre mice

  • Rubtsov et al. Regulatory T Cell-Derived Interleukin-10 Limits Inflammation at Environmental Interfaces. Immunity. 2008 Apr;28(4):546-58.

Group 3: Humanized Mice and Mutagenesis

  • Jordan et al. Complementation In Trans of Altered Thymocyte Development in Mice Expressing Mutant Forms of the Adaptor Molecule SLP76.  Immunity. 2008 Mar;28(3):359-69.  

Group 4: Reporter Mice:

  • Maynard et al. Regulatory T cells expressing interleukin 10 develop from Foxp3+ and Foxp3– precursor cells in the absence of interleukin 10. Nat Immunol. 2007 Sep;8(9):931-41. Epub 2007 Aug 12. 


DROP DEADLINE: Wednesday, April 6, 2011