Mitochondria from these Ppif knock-out mice exhibit an increased capacity to retain calcium and fail to swell/rupture in response to CaCl2. They are suitable for use in applications related to studies investigating oxidative stress- and ischemia–induced cell death.
Dr. Stanley J. Korsmeyer, Dana-Farber Cancer Institute
Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Brain architecture and cerebrovasculature are normal. No gene product (protein) is detected by immunoblot analysis of mitochondria isolated from liver tissue of mutant mice. Mitochondria from mutant mice have an increased capacity to retain calcium and fail to swell/rupture in response to CaCl2, suggesting abnormal permeability transition pore (PTP) function. Mouse embryonic fibroblasts (MEFs) derived from mutant mice are less susceptible to oxidative stress induced in vitro by exposure to hydrogen peroxide than MEFs derived from wildtype mice. In a model of ischemic brain injury employing a middle cerebral artery occlusion protocol, mutant mice exhibited a reduced infarct volume (37% in heterozygotes, and 62% in homozygotes) when compared to wildtype mice. This mutant mouse strain may be useful in studies investigating oxidative stress- and ischemia–induced cell death.
A targeting vector containing a loxP-flanked neomycin resistance gene cassette preceding exons 3-5 and a third loxP site downstream of exon 5 was electroporated into 129X1/SvJ-derived RW4 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a pMC-Cre gene to delete the neo gene (resulting in loxP-flanked exons 3-5 [CypDflox]) and then injected into C57BL/6 blastocysts. Chimeric mice were crossed to C57BL/6. The resulting heterozygotes were next crossed to FVB/N-Tg(EIIa-cre)C5379Lmgd/J (Stock No. 003314). Mice having been recombined such that exons 3-5 are replaced with a single loxP site (CypDnull) were intercrossed to generate mice homozygous for the deletion.
|Allele Name||targeted mutation 1.1, Michael A Moskowitz|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Ppif, peptidylprolyl isomerase F (cyclophilin F)|
|Strain of Origin||129X1/SvJ|
|Molecular Note||A targeting vector was designed to insert loxP sites around exons 3-5. Crossing with mice expressing cre in the germ line resulted in a null allele, as was demonstrated by immunoblot of liver samples.|
|Mutations Made By|| |
Dr. Stanley Korsmeyer, Dana-Farber Cancer Institute
When maintaining a live colony, these mice are bred as homozygotes.
When using the CyP-D KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #005740 in your Materials and Methods section.