Educational Level:

Roadmap Objectives:

• • Article: Brown CJ, Ballabio A, Rupert JL, Lafreniere RG, Grompe M, Tonlorenzi R, Wilard HF. (1991) A gene from the region of the human X inactivation centre is expressed exclusively from the inactive X chromosome. Nature 349 (6304):38-44
  • Content area/major concepts: Gene structure, transcription, and alternative splicing, DNA sequencing, Aneuploidy and Somatic cell hybrids
  • Methods or technology used to obtain data: DNA/RNA hybridization, cDNA library screening, gel electrophoresis, RNA slot and Northern blots, RT-PCR
  • How the CREATE strategy was used:
  • Biggest teaching challenge: Some previous knowledge of methods and/or papers assumed, primarily discusses molecular concepts but includes references to chromosomal abnormalities and approaches, aneuploidy and somatic cell hybrids, without discussion.
• • Article: Penny GD, Kay GF, Sheardown SA, Rastan S, Brockdorff N. (1996) Requirement for Xlist in x chromosome inactivation. Nature 379(6561):131-7
  • Content area/major concepts: Allelic variation, cell differentiation and gene expression, double-blind experiment
  • Methods or technology used to obtain data: cell culture, southern and RNA slot blots, restriction digest and RFLP, single nucleotide and primer extension assay (SNuPE), Targeted mutagenesis, Selectable markers and genetic screening, BrdU Labeling
  • How the CREATE strategy was used:
  • Biggest teaching challenge: Clear description of SNuPE assay may be difficult for students to find online. May wish to provide. Following cell strains and crosses somewhat complicated.
• • Article: Wutz, A and Jaenisch, R., (00) A shift from Reversible to Irreversible X inactivation is triggered during ES Cell Differentiation Molecular Cell. 5: 695-705 http://www.sciencedirect.com/science/article/pii/S1097276500802488
  • Content area/major concepts: Gene structure, splicing, short tandom repeats (STRs), chromatin modification
  • Methods or technology used to obtain data: Cell culture, genomic library screen, RT-PCR, northern blots, transgenic organisms, inducible gene expression, fluorescent in-situ hybridization (FISH), indirect immunofluorescence staining for histone acetylation
  • How the CREATE strategy was used:
  • Biggest teaching challenge: Long paper
• • Article: Kohlmaier, A., Savarese, F., Lachner, M., Martens, J., Jenuwein, T., Wutz, A. (04) A Chromosomal Memory Triggered by Xist Regulates Histone Methylation in X Inactivation. PLoS Biology. 2(7):0991-1003 http://biology.plosjournals.org
  • Content area/major concepts: Gene imprinting and chromatin modification, gene silencing, microsatellite markers, cell differentiation and transcription
  • Methods or technology used to obtain data: Indirect immunofluorescence staining for histone acetylation and methylation, fluorescent in-situ hybridization (FISH), chromatin immunoprecipitation, northern and western blots, deletion analysis, gene induction and timed expression
  • How the CREATE strategy was used:
  • Biggest teaching challenge: Long paper

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