These Adar knockout mice die between E11.5 and E12.5 due to a severe liver defect. Hematopoietic tissues including erythroid and myeloid/granuloid progenitors are significantly reduced in the yolk sac, fetal liver and peripheral blood. This mutant mouse strain may be useful in studies of RNA editing and hematopoiesis.
Peter Seeburg, Max Planck Institute
Kazuko Nishikura, The Wistar Institute
Homozygotes: Homozygous embryos die between E11.5 and E12.5 due to a severe liver defect. Expected Mendelian numbers of embryos are recovered at E11.5 and appear normal, however, by E12, embryos exhibit a significant reduction in liver size, a progressively pale yolk sac and developmental retardation. By E12.5, all embryos are dead. By E11.5, fetal livers have few hematopoietic cells, an accumulation of blood in enlarged live spaces, many cells with pyknotic nuclei and few erythroid cells. Hematopoietic tissues including erythroid and myeloid/granuloid progenitors are significantly reduced in the yolk sac, fetal liver and peripheral blood. This mutant mouse strain may be useful in studies of RNA editing and hematopoiesis.
Heterozygote: Mice that are heterozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.
A targeting vector containing a loxP-flanked PGKneo cassette was placed in intron 6 and an additional loxP was placed in intron 9. The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1 derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred to mice carrying Tg(CMV-cre)1Cgn, the cre deleter transgene, consequently removing the neomycin cassette and exons 7-9 and leaving a loxP site upstream of exon 10. The strain was transferred to Dr. Nishikura at the Wistar Institute and backcrossed to C57BL/6 (see SNP note below) for at least 10 generations. During backcrossing the Cre transgene was bred out of the line. Upon arrival, mice were bred to C57BL/6J for at least 1 generation to establish the colony.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, all 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6N genetic background.
|Allele Name||targeted mutation 1.1, Peter H Seeburg|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||Adarf; adarf7-9|
|Gene Symbol and Name||Adar, adenosine deaminase, RNA-specific|
|Strain of Origin||(129X1/SvJ x 129S1/Sv)F1-Kitl+|
|Molecular Note||A floxed neo selection cassette was placed upstream of exon 7 in the SmaI restriction site and an additional loxP site was inserted downstream of exon 9. Correct targeting was confirmed by genomic PCR. LoxP-flanked (floxed) sequence encoded residues 707?870 of the interferon-inducible form of ADAR1 (GenBankTM/EBI accession number AAK16102), including essential regions for editing activity.|
|Mutations Made By|| |
Peter Seeburg, Max Planck Institute
While maintaining a live colony, these mice are bred as heterozygotes. Mice homozygous for the mutation are not viable.
When using the B6.129(Cg)-Adartm1.1Phs/KnkMmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #34620 in your Materials and Methods section.