Because loss-of-function experiments in medaka and zebrafish had relied on morpholino, whose effect is transient and lasts only a few days post fertilization, developmental biology has been the major target of research using these small laboratory fish. With the advent of targeting-induced local lesions in genome (TILLING) technology, the teleost has reemerged as a model animal in which the specific genes can be disrupted. The potential research field is now expanded to slow and time-consuming biological phenomena such as carcinogenesis, aging, and degeneration. p53 is a multifunctional transcription factor involved in such various biological processes. Among other vertebrates, medaka is especially suited for the TILLING approach because of the compact genome and inbred strains, which makes amplicon design and screening fast and efficient. We have generated several lines of p53 mutant medaka, including two nonsense mutants. These lines showed the impaired transcriptional regulation of the cell cycle and apoptosis and the increased incidence of spontaneous tumorigenesis. The mutant medaka isolated by TILLING will occupy an important and valuable position in various research fields in the future.
KeywordsAcridine Orange Morpholino Antisense Oligonucleotide Median Lifespan Target Gene Induction Medaka Genome
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