Viability E14.5-E15.5 Secondary Screen IMPC_EVO_001

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Purpose

To assess the viability, sub-viability, and lethality of homozygous embryos at E14.5 or E15.5

Experimental Design

  • Set up timed matings with heterozygous mice
  • Day 0 is defined as the midpoint of the prior dark cycle following the identification of a copulation plug.
  • Collect embryos at E14.5 or E15.5
  • Collect tissue and genotype embryos.

Procedure

  1. Set up timed mating with heterozygous animals. Aim to dissect and collect >=28 alive embryos, otherwise lethal and subviable calls cannot be made. If more than three homozygous pups are produced before 28 pups are genotyped, a viable call can be made.
  2. Collect tissue for genotyping and (OPTIONAL) score Gross Morphology and/or process for Histopathology and or Imaging.
  3. Genotype all embryos and
    1. Strains that produce NO existing homozygous embryos will be considered LETHAL (complete embryonic lethality [MP:TBC]).
    2. Strains that produce NO live (absence of heartbeat) homozygous embryos will be considered LETHAL (complete embryonic lethality [MP:TBC]).
    3. Strains that produce live homozygous embryos but with an obvious defect will be left to the discretion of the center with the decision and reason recorded in the parameters.
    4. X-linked strains that produce NO live hemizygous male embryos from female carriers will be considered LETHAL (complete embryonic lethality [MP:TBC]).
  4. Flag strains that produce less than normal numbers of homozygous/hemizygous male progeny
    1. Strains that produce <50% expected homozygous progeny will be annotated as partial embryonic lethality [MP:TBC].
    2. X-linked strains that produce <50% expected male hemizygous progeny from female carriers will be considered partial embryonic lethality [MP:TBC].

Notes

Data QC

All genotypes should be collected using validated assays.

Y chromosome assay required for X-linked lethal strains.

 

Data Analysis, annotation and display (+statistics)

 

Total Embryos: All, WT, Het, Hom
    •Alive, dead, and defect (all genotyped)
Total Dead: All, WT, Het, Hom

Total Defect (Alive or Dead):  All, WT, Het, Hom
    •Abnormal and dead embryos
Litter size: all genotyped embryos
    •ignore partials and reabsorptions.

Parameters & Metadata

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Name Version Type Req. Upload Req. Analysis Annotation Increment Options Ontology Options Unit Data Type Derived History
Name Version Type Req. Upload Req. Analysis Annotation Increment Option Ontology Options Unit Data Type Derived History