Implementation of the modified-SHIRPA protocol for screening of dominant phenotypes in a large-scale ENU mutagenesis program |
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Authors: | Hiroshi Masuya Maki Inoue Yumiko Wada Aya Shimizu Junko Nagano Akiko Kawai Ayako Inoue Tomoko Kagami Taeko Hirayama Ayako Yamaga Hideki Kaneda Kimio Kobayashi Osamu Minowa Ikuo Miura Yoichi Gondo Tetsuo Noda Shigeharu Wakana Toshihiko Shiroishi |
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Institution: | (1) Functional Genomics Research Group, RIKEN Genomic Sciences Center, 3-1-1 Kouyadai, Tsukubai Ibaraki, 305-0074, Japan;(2) Population and Quantitative Genomics Team, RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama City Kanagawa, 230-0045, Japan |
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Abstract: | SHIRPA is a three-stage protocol for the comprehensive assessment of primarily mouse behavior. The first stage consists of
high-throughput phenotyping of 33 behavioral observations and 7 metabolic or disease observations. We modified this part of
the protocol by integrating new morphologic observations into the initial phenotype assay of behavior and dysmorphology. Behavioral
observations assessed by this protocol, now referred to as the “modified-SHIRPA,” are compatible with the original “SHIRPA”
protocol. Using modified-SHIRPA, we screened dominant phenotypes of more than 10,000 G1 progeny generated by crossing DBA/2J females with ENU-treated C57BL/6J males. To date, we have obtained 136 hereditary-confirmed
mutants that exhibit behavioral and morphologic defects. Some independent mutant lines exhibited similar phenotypes, suggesting
that they may represent alleles of the same gene or mutations in the same genetic pathway. They could hold great potential
for the unraveling of the molecular mechanisms of certain phenotypes. |
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