Guide to Techniques in Mouse Development, Part B: Part B

Covers different techniques for the cryopreservation of gametes and embryos, production of transgenic and null (knockout) animals (use of ES cells), generation of conditional/inducible mutant animals, use of gene-trap mutagenesis, analysis of allele-specific expression, use of new reporter constructs, humanizing of transgenic animals, and more.

Section I. Transgenesis 1. Lentivirus transgenesis 2. Germline modification using mouse spermatogonial stem cells 3. Embryonic in vivo electroporation in the mouse Section II. Transposons 4. Current applications of transposons in mouse genetics 5. Functional genomics in the mouse using the Sleeping Beauty transposon system 6. The use of DNA transposons for cancer gene discovery in mice Section III. Recombinases 7. A practical summary of site specific recombination, conditional mutagenesis, and tamoxifen induction of CreERT2 8. A recombineering pipeline to make conditional targeting constructs 9. Confirmation of recombination site functionality in gene targeting vectors using recombinase-expressing bacteria 10.Genetic fate mapping using site-specific recombinases 11. Mapping cell fate and function using recombinase-based intersectional strategies Section IV. Mutagenesis 12. Genome-wide forward genetic screens in mouse ES cells 13. Gene trap mutagenesis in the mouse 14. A wider context for gene trap mutagenesis 15. Mouse mutagenesis with the chemical supermutagen ENU 16. Phenotype-driven mouse ENU mutagenesis screens 17. Using ENU mutagenesis for phenotype-driven analysis of the mouse Section V. Gene Knockdowns 18. Exploration of self-renewal and pluripotency in ES cells using RNAi 19. Transgenic RNAi applications in the mouse 20. Gene knockdown in the mouse through RNAi 21. In vivo analysis of gene knock-down in tetracycline-inducible shRNA mice 22. The power of reversibility: Regulating gene activities via tetracycline controlled transcription Section VI. Gene Expression Profiling 23. Gene expression profiling of mouse oocytes and preimplantation embryos 24. Interrogating the transcriptome of oocytes and preimplantation embryos 25. Gene expression profiling of mouse embryos with microarrays