These R1.40 transgenic mice express all mRNA and protein isoforms of the human amyloid beta (A4) precursor protein APP containing the Familial Alzheimer's Disease (FAD) Swedish mutation K670N/M671L, and may be useful in studying the pathogenesis of Familial Alzheimer's Disease and possible therapeutic treatments.
Bruce Lamb, The Cleveland Clinic Foundation
These R1.40 transgenic mice express all mRNA and protein isoforms of the human amyloid beta (A4) precursor protein APP containing the Familial Alzheimer's Disease (FAD) Swedish mutation K670N/M671L. Transgene expression (mRNA and full-length protein) is 2 to 3 fold the wild-type mouse App expression level in the hemizygous state in brain tissue as revealed by RT-PCR and Western Blot analysis. Transgene expression pattern mimics wild-type mouse gene expression patterns. 3 to 4 month old transgenic mice, maintained on a C57BL/6J background, exhibit spontaneous seizure-like events (abnormal spiking) in EEG readings, without abnormal behavior and are more susceptible to kainic acid induced seizures (Vogt et al. Neurobiol Aging 2009).These R1.40 transgenic mice may be useful in studying the pathogenesis of Familial Alzheimer's Disease and possible therapeutic treatments.
Of note, this is one of two 129S1/SvImJ congenic R1.40 transgenic strains (129S1-R1.40) segregating for transgene copy number; one with lower trangene copy number (129S1.129(Cg)-Tg(APPSw)40Btla/Mmjax) and one with higher transgene copy number. No phenotype differences between the low and high copy number strains were reported, and the high copy number strain was discarded (June 2010).
In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype could vary from that originally described. A publication (Lehman et al. 2003 Hum Mol Genet 12:2949) compares the well characterized B6-R1.40 strain (B6.129-Tg(APPSw)40Btla/Mmjax) with two additional congenic strains, D2-R1.40 (D2.129(B6)-Tg(APPSw)40Btla/Mmjax) and 129S1-R1.40 (129S1.129(Cg)-Tg(APPSw)40Btla/Mmjax). While these three congenic strains have similar levels of holo-APP in brain tissue, the levels of brain APP C-terminal fragments (CTFs) vary depending upon genetic background. Brain and plasma levels of amyloid beta-40 and -42 are variable as well (B6-R1.40 > 129S1-R1.40 > D2-R1.40). In addition, the congenic strains exhibited dramatic alterations in the age of onset of amyloid beta deposition; in contrast to 14 month old homozygous B6-R1.40 mice, homozygous D2-R1.40 and 129S1-R1.40 mice do not develop amyloid beta deposits in the parietal or frontal cortex even by 20 months of age. The donating investigator further reports that the A-R1.40 strain (A.129(B6)-Tg(APPSw)40Btla/Mmjax) exhibits levels of amyloid beta comparable to the B6-R1.40 strain, but with later onset. Therefore, APP processing and amyloid beta metabolism and deposition are modified by the genetic background. While the 129S1-R1.40 strain can be easily maintained as homozygotes, the donating investigator reports increased mortality in young homozygotes on the other genetic backgrounds, with D2-R1.40 and A-R1.40 more severely affected than B6-R1.40.
A 650 kb YAC transgene containing the entire human amyloid beta (A4) precursor protein (APP) gene, and approximately 250 kb of flanking sequence, was altered to include the Swiss mutation K670N/M671L associated with Familial Alzheimer's Disease (FAD). This transgene was injected into (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells. Founder animals (line R1.40) were established and bred to C57BL/6J for an unknown number of generations. The mice were next bred to DBA/2J for seven generations. After this, transgenic mice were bred to 129S1/SvImJ for at least 24 generations to generate this 129S1-R1.40 transgenic strain prior to arrival at the MMRRC at The Jackson Laboratory. This transgene inserted on Chromosome 13.
After approximately 5 generations of breeding 129S1-R1.40 transgenic mice together, two populations of mice were identified; one with lower transgene copy number (129S1.129(Cg)-Tg(APPSw)40Btla/Mmjax) and one with higher copy number. No phenotype differences between the low and high copy number strains were reported, and the high copy number strain was discarded (June 2010).
|Expressed Gene||APP, amyloid beta precursor protein, human|
|Site of Expression|
|Allele Name||transgene insertion 40, Bruce Lamb|
|Allele Type||Transgenic (Humanized sequence, Inserted expressed sequence)|
|Allele Synonym(s)||APPK670/M671; APPK670N/M671L; R1.40; R1.40-YAC; Tg(APP*Swe)40Btla|
|Gene Symbol and Name||Tg(APPSw)40Btla, transgene insertion 40, Bruce Lamb|
|Gene Synonym(s)||APPK670/M671; APPK670N/M671L; R1.40; R1.40-YAC|
|Promoter||APP, amyloid beta precursor protein, human|
|Expressed Gene||APP, amyloid beta precursor protein, human|
|Strain of Origin||(129X1/SvJ x 129S1/Sv)F1-Kitl<+>|
|Molecular Note||The transgenic insertion comprises four to eight copies of a 650-kb YAC containing the entire 400-kb human amyloid precursor protein gene with the Swiss familial Alzheimer disease (FAD) mutation and approximately 250 kb of flanking human DNA. The double mutation (K670N/M671N) comprises a G-to-A transition converting the lysine codon at position 670 to an asparagine codon and an A-to-C transversion replacing the methionine at amino acid 671 with leucine. RT-PCR and western blot analysis revealed that hemizygous transgenic mice express all major human APP mRNA and protein isoforms in brain, in parallel with the corresponding mouse versions, at approximately three times the levels of the latter. Total brain levels of the Alzheimer disease- associated 42-amino acid amyloid-beta peptide detected by ELISA are 7-8-fold and 15-20-fold higher, respectively, in mice hemizygous and homozygous for the mutant gene than in hemizygotes for the wildtype human gene. Levels of a cell-associated, beta-secretase-generated 13.5 kDa C-terminal peptide containing the amyloid beta domain and of soluble amyloid beta peptides are significantly elevated in brains of mice with the mutant, versus wildtype, transgene, while alpha-secretase products are diminished.|
|Mutations Made By|| |
Bruce Lamb, The Cleveland Clinic Foundation
When maintaining a live colony, homozygous mice may be bred together. While increased mortality is observed in young homozygotes on the other genetic backgrounds, the donating investigator reports that this strain can be easily maintained as homozygous.
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