Systematic Approach to Evaluation of Mouse Mutations includes information on medical photography, grafting procedures, how to map the genes and evaluate the special biological characteristics of the mice. Read more Read less. Review "The Jackson Lab crew, with assists from other contributors, have put together a reference for scientists, mouse geneticists, managers and lab animal veterinarians. Sundberg, Dawnalyn Boggess. No customer reviews. Share your thoughts with other customers. Write a customer review. Discover the best of shopping and entertainment with Amazon Prime.
In this case, the dead male was stored in a sealed plastic bag to prevent dessication in the refrigerator until the IVF. Another time we rescued a mouse strain from a single spontaneous male mutant with ectopic caudal limbs in the place of a penis polypodia mutation — Ppd gene on the X chromosome. In this case you can send the old females to the Jackson Laboratory for ovarian transplantation into a immunocompromised female to prevent graft rejection and you may be able to recover the line this way.
The Transgenic Core does not offer this service. If you took the precaution of cryopreserving the line as frozen eggs, the Core can thaw out the eggs and recover the line.
Systematic Approach to Evaluation of Mouse Mutations - Google Books
If sperm were cryopreserved we may or may not be able to recover the line by IVF or ICSI, depending on the genetic background of the sperm. We recommend that frozen sperm be tested for successful IVF before a line is discontinued. If the line was derived from an ES cell clone, it can be re-injected to generate a new set of germline chimeras.
Three transgene Tg transmission patterns occur in transgenic founder animals. In these cases it is advisable to verify the transgenic status of the founder and to breed the mouse as efficiently as possible. Different insertion sites usually segregate independently. Southern blot analysis of these offspring is used to group mice have according to insertion site. This simplifies analysis of transgene expression since expression patterns associated each independent insertion are isolated from each other. Offspring of transgenic founders transmit the Tg as normal Mendelian gene, regardless of whether the founder was mosaic or multi-integrant.
Two transmission patterns occur in embryonic stem ES cell-mouse chimeras.
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Infertile male chimeras may also result. Since the ES cells are XY only male chimeras should be bred.
Mutation discovery in mice by whole exome sequencing
We recommend that you take a male chimera through six litters before deciding to give up on it. If a chimera has a high transmission ratio, you should consider mating it directly with females from the mouse strain that matches the origin of the ES cell line to obtain your targeted gene mutation on an inbred background. We recommend that transgenic lines be maintained as hemizygotes. The primary disadvantage of maintaining lines as hemizygotes is that all offspring need to be genotyped to determine which are transgenic. Establishment of homozygous transgenic lines is is costly and entails additional breeding to produce the first homozygotes.
Additional test breeding is needed to ensure homozygosity. Additional time is required to establish a homozygous line from a mosaic founder with limited transgene transmission. Extra breeding will be necessary to establish homozygous lines for each integration site in founders with multiple integrations. If this is not done carefully, animals with combinations of different integration sites will result. This will complicate interpretation of experimental results since different integration sites can have different transgene expression patterns.
Trends Genet. Affected mice may display a phenotype unassociated with the transgene which may obscure the effects of transgene expression. Unequal crossing-over between transgene arrays in homozygotes may result in transgene rearrangement which will affect expression. This kind of instability occurs less often in hemizygotes.
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Many other difficulties can be avoided by maintaining hemizygous transgenic lines. The status of the gene map of the human chromosomes.
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Woodworth-Gutai, C. Kane-Kass, J. Houston, J. Heath, and N. Most highly repeated dispersed DNA families in the mouse genome.
- Systematic Approach to Evaluation of Mouse Mutations - CRC Press Book.
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These may vary with age of mice, parity, and environmental conditions e. Hogan, R. Beddington, F. Constantini, and E. Lacey, published by Cold Spring Harbor Laboratory. This book has an excellent bibliography for information about mouse strains and breeding. Some investigators will need to know the alleles at various polymorphic loci carried by the transgenic founders.
This information can be found by looking up the genotypes of the mice used to obtain fertilized eggs for microinjection in:. Genetic Variants and Strains of the Laboratory Mouse, eds. Lyon and A. Roderick and J. Guidi, published by the Oxford University Press. Search for:.