These PWS-ICflox6kb mice may be useful in generating conditional mutations for studying the role of Snrpn and the temporal and developmental requirements of the Prader-Willi imprinting center.
James Resnick, University of Florida
Mice homozygous for this PWS-ICflox6kb allele are viable and fertile, with loxP sites flanking the 6 kb Prader-Willi Syndrome imprinting center (PWS-IC) region around exon 1 of the Snrpn gene . When bred to mice that express Cre recombinase, the resulting offspring will have this area deleted in the cre-expressing tissue(s). Because Snrpn is imprinted and only expressed from the paternal allele, breeding PWS-ICflox6kb males with cre-expressing females is required to generate deleted offspring with the knockout phenotype. These PWS-ICflox6kb mice be useful in studying the temporal and spatial functions of the Prader-Willi imprinting center.
When crossed to female mice carrying the Tg(CMV-cre)1Cgn allele (see Stock No. 006054), widespread Cre recombinase expression prior to implantation results in postnatal lethality.
When crossed to female mice carrying the Tg(Nes-cre)1Kln allele (see Stock No.003771), Cre recombinase expression in brain precursors cells starting at E10.5 results in a growth retardation, but is not lethal.
A targeting vector was designed to insert a loxP site upstream of exon 1 followed by a loxP-flanked neomycin resistance (neo) cassette downstream of exon 1. The construct was electroporated into 129S1/Sv-derived CJ7 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a cre-expressing plasmid to remove the neo selection cassette, but retain the loxP-flanked 6 kb Prader-Willi Syndrome-imprinting center (PWS-IC) region centered around exon 1. Chimeric mice crossed to C57BL/6J and offspring were backcrossed to C57BL/6J for 11 generations.
Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
|Allele Name||targeted mutation 2.1, Karen A Johnstone|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Snrpn, small nuclear ribonucleoprotein N|
|Strain of Origin||129S1/Sv-Oca2+ Tyr+ Kitl+|
|Molecular Note||A loxP site was inserted 3.7 kb upstream of exon 1. A floxed neo cassette was inserted downstream of exon 1. Cre-mediated recombination first removed the neo cassette and left the 6 kb encompassing the Prader-Willi syndrome imprinting center (PWS-IC) floxed.|
|Mutations Made By|| |
James Resnick, University of Florida
While maintaining a live colony, these mice are bred as homozygotes.
Because Snrpn is imprinted and only expressed from the paternal allele, breeding PWS-ICflox6kb males with cre-expressing females is required to generate offspring with the deleted flanked sequence and the knockout phenotype.
When using the B6.129S1-Snrpntm2.1Kaj/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #018395 in your Materials and Methods section.
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