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Mouse Phenotypes: Generation and Analysis of Mutants, Second Edition: A Laboratory Manual


Subject Area(s):  Developmental BiologyBiotechnologyGenetics

By Virginia E. Papaioannou; Richard R. Behringer

Download a Free Excerpt from Mouse Phenotypes: Generation and Analysis of Mutants, Second Edition: A Laboratory Manual:

Preface
Chapter 3. Mouse Gene-Targeting Strategies for Maximum Ease and Versatility
Index


© 2024 • 296 pages, illustrated (78 color and 60 B&W), index
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ISBN  978-1-621824-18-3
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  •     Description    
  •     Contents    

Description

In this postgenome era, the large number of mouse mutations available is revolutionizing biomedical research. Large-scale mutagenesis projects have resulted in the modification of thousands of mouse genes and, more recently, with CRISPR–Cas gene editing, generation of custom mutations is easier than ever. The interpretation of mutant phenotypes is critically important for understanding how changes in the genome result in changes in the phenotype. Complex phenotypes resulting from gene manipulation or spontaneous mutation, however, provide a huge analytic challenge. Where does one start with the analysis? How does one dissect the phenotype and begin to understand the molecular and developmental consequences of the genotype? How does one make the best use of the valuable resource that you have developed?

Mouse Phenotypes: A Handbook of Mutation Analysis, Second Edition, provides a road map for such analysis. Advice and practical guidance are provided, starting with the planning stages of gene targeting, CRISPR–Cas gene editing, or mutagenesis experiments, through the complete analysis of complex phenotypes, including tips on breeding strategies, studying dominant mutations, and characterizing a mutant mouse with no apparent phenotypic abnormalities. A systematic approach to the analysis, complemented with simple protocols, will help users make a comprehensive assessment of a particular phenotype resulting from a mutation and determine the timing and likely causes of specific developmental failures. Investigators using the laboratory mouse in genetic studies designed to understand mammalian biology will benefit from the authors’ experience and advice in analyzing mutant phenotypes.

Contents

Preface
Chapter 1. Strategies for the Production and Phenotypic Analysis of Mutations in the Mouse
Chapter 2. Obtaining or Generating Gene Mutations in Mice
Chapter 3. Mouse Gene-Targeting Strategies for Maximum Ease and Versatility
Chapter 4. Embryonic Stem Cell Gene Targeting and Chimera Production in Mice
PROTOCOLS
Protocol 4.1 Counting Murine Corpora Lutea to Determine the Number of Oocytes Ovulated
Protocol 4.2 Cross-Fostering of Newborn Mice to Counteract Bad Parenting
Chapter 5. Recovering a Targeted Mutation in Mice from Embryonic Stem Cell Chimeras or CRISPRCas Founders
PROTOCOL
Protocol 5.1 Sex Genotyping Mice by Polymerase Chain Reaction
Chapter 6. Strategies for Maintaining Mouse Mutations
PROTOCOL
Protocol 6.1 Backcrossing to Generate a Congenic Mouse Strain
Chapter 7. Special Breeding Techniques for Use in Mouse Mutation Analysis
Chapter 8. Phenotypic Analysis: Assessing Timing of Recessive Prenatal Lethality in Mice
PROTOCOL
Protocol 8.1 Establishing Control Frequency of Embryonic and Fetal Loss in a Mutant Mouse Colony
Chapter 9. Phenotypic Analysis of Preimplantation Lethality in Mice
PROTOCOLS
Protocol 9.1 Phenotypic Analysis of Preimplantation Mouse Embryos in Culture
Protocol 9.2 Cell Counting Techniques for Preimplantation Mouse Embryos Using Fluorescent DNA Dyes
Chapter 10. Phenotypic Analysis of Periimplantation to Mid-Gestation Lethality in Mic
PROTOCOL
Protocol 10.1 Orientation of Implanted or Dissected Mouse Embryos for Histological Sectioning
Chapter 11. Analysis of Mid- to Late-Gestation Phenotypes in Mice
Chapter 12. Uncovering Phenotypes in Mutant Mice by Determining Embryo, Organ, Tissue, and Cell Developmental Potential
Chapter 13. Phenotypic Analysis of Perinatal Lethality in Mice
Chapter 14. Analysis of Postnatal Mutant Phenotypes in Mice
Chapter 15. The No Phenotype Challenge in Analyzing Mutant Mice
PROTOCOL
Protocol 15.1 Necropsy Guide for the Collection of Tissues from Mice with or without Tumors
Chapter 16. Phenotypic Analysis of Dominant Mutant Effects in Mice
PROTOCOL
Protocol 16.1 Isolation of XO Subclones from XY Murine Embryonic Stem Cells
Chapter 17. Getting around an Early Lethal Phenotype in Mice with Chimeras
Chapter 18. Tissue- and/or Temporal-Specific Mutations in Mice Using Conditional Alleles
APPENDIXES
1. Useful Books and Other Resources
2. Examples of Phenotype Analysis
3. Glossary
4. General Safety and Hazardous Material Information
Index