Genomic Imprinting

1. Front Matter

   Pages i-xii

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2. Generation of Monoparental Embryos for Investigation into Genomic Imprinting

   • Wendy L. Dean, Gavin Kelsey, Wolf Reik

   Pages 1-19

3. Deriving and Propagating Mouse Embryonic Stem Cell Lines for Studying Genomic Imprinting

   • Jeffrey R. Mann

   Pages 21-39

4. Balanced Translocations for the Analysis of Imprinted Regions of the Mouse Genome

   • Anne C. Ferguson-Smith, Maxine Tevendale, Pantelis Georgiades, Valerie Grandjean

   Pages 41-54

5. Production of YAC Transgenic Mice by Pronuclear Injection

   • Justin F.-X. Ainscough, Rosalind M. John, Sheila C. Barton

   Pages 55-65

6. A Transgenic Approach to Studying Imprinted Genes

   • Rosalind M. John, Justin F.-X. Ainscough, Sheila C. Barton

   Pages 67-81

7. Methylation-Sensitive Genome Scanning

   • Izuho Hatada, Tsunehiro Mukai

   Pages 83-100

8. Subtraction-Hybridization Method for the Identification of Imprinted Genes

   • Fumitoshi Ishino, Yoshimi Kuroiwa, Naoki Miyoshi, Shin Kobayashi, Takashi Kohda, Tomoko Kaneko-Ishino

   Pages 101-112

9. Identification of Imprinted Loci by Methylation

   • Rachel J. Smith, Gavin Kelsey

   Pages 113-132

10. Ribonuclease Protection

    • Joanne L. Thorvaldsen, Marisa S. Bartolomei

    Pages 133-144

11. Quantitative RT-PCR-Based Analysis of Allele-Specific Gene Expression

    • Judith Singer-Sam, Chunguang Gao

    Pages 145-152

12. Allele-Specific In Situ Hybridization (ASISH)

    • Rolf Ohlsson, Kristian Svensson, Hengmi Cui, Helena Malmikumpu, Gail Adam

    Pages 153-167

13. RNA-FISH to Analyze Allele-Specific Expression

    • Giovanna Braidotti

    Pages 169-180

14. Flow Cytometry and FISH to Investigate Allele-Specific Replication Timing and Homologous Association of Imprinted Chromosomes

    • Janine LaSalle, Marc Lalande

    Pages 181-192

15. Southern Analysis Using Methyl-Sensitive Restriction Enzymes

    • Tom Moore

    Pages 193-203

16. A PCR-Based Method for Studying DNA Methylation

    • Mira Ariel

    Pages 205-216

17. Bisulfite-Based Methylation Analysis of Imprinted Genes

    • Sabine Engemann, Osman El-Maarri, Petra Hajkova, Joachim Oswald, Joern Walter

    Pages 217-228

18. Direct Analysis of Chromosome Methylation

    • Déborah Bourc’his, Evani Viegas-Péquignot

    Pages 229-242

19. In Vitro Methylation of Predetermined Regions in Recombinant DNA Constructs

    • Ilse Van den Wyngaert, Roger L. P. Adams, Stefan U. Kass

    Pages 243-250

20. In Vitro Methylation of Specific Regions in Recombinant DNA Constructs by Excision and Religation

    • Ghislaine Dell, Marika Charalambous, Andrew Ward

    Pages 251-258

Genomic imprinting is the process by which gene activity is regulated according to parent of origin. Usually, this means that either the maternally inherited or the paternally inherited allele of a gene is expressed while the opposite allele is repressed. The phenomenon is largely restricted to mammals and flowering plants and was first recognized at the level of whole genomes. Nuclear transplantation experiments carried out in mice in the late 1970s established the non-equivalence of the maternal and paternal genomes in mammals, and a similar conclusion was drawn from studies of interploidy crosses of flowering plants that extend back to at least the 1930s. Further mouse genetic studies, involving animals carrying balanced translocations (reviewed in Chapter 3), indicated that imprinted genes were likely to be widely scattered and would form a minority within the mammalian genome. The first imprinted genes were identified in the early 1990s; over forty are now known in mammals and the list continues steadily to expand.

"...a multi-authored volume in which the methods of gene imprinting are discussed in detail in 25 chapters. The protocols are very detailed with multiple schemes and figures." - Journal of Pediatric Endocrinology and Metabolism

• Department of Biology and Biochemistry, University of Bath, Claverton Down, UK

  Andrew Ward

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