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

ISBN-13: 9780123848826 / Angielski / Miękka / 2010 / 580 str.

Paul Wassarman

Opis

Guide to Techniques in Mouse Development, Part B, is an authoritative guide to different methods used in enzymology, focusing on investigating mouse development using technological advances. The text provides information regarding the principles of the methods in mouse development, and it offers readers reliable experimental protocols and recipes described comprehensively by leaders in the field of enzymology. The text is divided into three sections and organized into 25 chapters. Below are several concepts covered by the text:

• Lentivirus transgenesis o Germline modification using mouse stem cells
• Electroporation
• Applications of transposons in mouse genetics
• Functional genomics using transposon systems
• The use of DNA transposons in detecting cancer genes in mice
• Recombination, conditional mutagenesis and induction of tamoxifen
• Genetic fate mapping using recombinases
• Genetic screens mouse ES cells
• Gene trap mutagenesis
• Mouse mutagenesis
• Self- renewal and pluripotency
• Transgenic RNAi applications
• Gene knockdowns
• Tetracycline-controlled transcription
• Gene expression profiling of mouse embryos

The book is a comprehensive guide for students and professionals in genetics, cytology and molecular biology, who will find this book invaluable for their learning and practice.

• Update of volume 225, Guide to Techniques in Mouse Development, edited by P.M. Wassarman and M.L. DePamphilis and published in 1993.
• Comprehensively covers:
• new 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
• transcript profiling of mouse development
• imaging of mouse development
• rederivation of animals and use of mouse genomics

Spis treści

Section I. Transgenesis1. Lentivirus transgenesis2. Germline modification using mouse spermatogonial stem cells3. Embryonic in vivo electroporation in the mouse

Section II. Transposons4. Current applications of transposons in mouse genetics5. Functional genomics in the mouse using the Sleeping Beauty transposon system6. The use of DNA transposons for cancer gene discovery in mice

Section III. Recombinases7. A practical summary of site specific recombination, conditional mutagenesis, and tamoxifen induction of CreERT28. A recombineering pipeline to make conditional targeting constructs 9. Confirmation of recombination site functionality in gene targeting vectors using recombinase-expressing bacteria10.Genetic fate mapping using site-specific recombinases11. Mapping cell fate and function using recombinase-based intersectional strategies

Section IV. Mutagenesis12. Genome-wide forward genetic screens in mouse ES cells13. Gene trap mutagenesis in the mouse14. A wider context for gene trap mutagenesis 15. Mouse mutagenesis with the chemical supermutagen ENU16. Phenotype-driven mouse ENU mutagenesis screens17. Using ENU mutagenesis for phenotype-driven analysis of the mouse

Section V. Gene Knockdowns18. Exploration of self-renewal and pluripotency in ES cells using RNAi19. Transgenic RNAi applications in the mouse 20. Gene knockdown in the mouse through RNAi21. In vivo analysis of gene knock-down in tetracycline-inducible shRNA mice22. The power of reversibility: Regulating gene activities via tetracycline controlled transcription

Section VI. Gene Expression Profiling23. Gene expression profiling of mouse oocytes and preimplantation embryos24. Interrogating the transcriptome of oocytes and preimplantation embryos25. Gene expression profiling of mouse embryos with microarrays

O autorze

Philippe Soriano,is Professor of Cell, Developmental & Regenerative Biology and Professor of Oncological Sciences, Icahn School of Medicine at Mount Sinai, NY, USA