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In publishing the research results to be obtained by use of the BIOLOGICAL RESOURCE, an acknowledgment to the DEPOSITOR is requested.
The recipient is requested to cite the literature designated by the DEPOSITOR.
Sci Rep. 2012;2:945.
Kawamata M, Ochiya T.
Two distinct knockout approaches highlight a critical role for p53 in rat.
Gene Affected
p53遺伝子を欠損。CAG promoter, AmCyan1, IRES, neo, pA 遺伝子がノックインにより組み込まれている。
Origin
LEA rats were provided fron Kyoto Bioresource Center in 2006. Then LEA rat ES cells having Oct4-Venus gene and Oct4-Venus Tg rat strain (LEA-Tg(Pou5f1-YFP*)3Ncco) were established.
ES cells were establised from Oct4-Venus Tg rat, p53 heterozygous-ES cells were made by gene targeting method and finally p53-heterozygous rat strain (LEA-Tp53tm1(AmCyan1)Ncco) was established by germline transmission. Deposited p53-heterozygous rats do not have Oct4-Venus.
Vector: AmCyan1 (Clontech)
Strain characteristics
AmCyan1 expression under CAG-promoter and deletion of p53 gene expression. Almost all p53 heterozygous rats die of tumor within one year of age.
Breeding Conditions
mating between heterozygous rats
Genotyping
PCR analysis
wild type PCR product: 207 bp product
Forward: GGAGGGAAGGGTATCACACA
Reverse: CACCAGCTCACCAAGGCCTA
Mutant PCR product: about 1100 bp
Forward: ATTCGCAGCGCATCGCCTTCT
Reverse: GCTGGAGATATGGGTGGGTC
KOD version2 (TOYOBO)
94℃: 2 min, 35 cycles (98℃: 20 sec, 62℃: 20 sec, 68℃: 1 min), 68℃: 5 min, 4℃
References
Kawamata M, Ochiya T. Generation of genetically modified rats from embryonic stem cells. Proc Natl Acad Sci U S A. 2010 Aug 10;107(32):14223-8.
Kawamata M, Ochiya T. Two distinct knockout approaches highlight a critical role for p53 in rat development. Sci Rep 2012 2: 945