Implementation of the modified-SHIRPA protocol for screening of dominant phenotypes in a large-scale ENU mutagenesis program
Implementation of the modified-SHIRPA protocol for screening of dominant phenotypes in a large-scale ENU mutagenesis program
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DOI：
10.1007/s00335-005-2430-8
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发表时间：
2005-11-01
期刊：
影响因子：
2.5
通讯作者：
Shiroishi, T
中科院分区：
文献类型：
--
作者：
Masuya, H;Inoue, M;Shiroishi, T
SHIRPA是一个三阶段的协议，主要用于小鼠行为的综合评估。第一阶段包括33项行为观察和7项代谢或疾病观察的高通量表型。我们通过将新的形态学观察整合到行为和畸形的初始表型分析中来修改这部分协议。通过该协议评估的行为观察，现在被称为“修改后的SHIRPA”，与原始的“SHIRPA”协议兼容。利用改进的shirpa，我们筛选了DBA/2J雌性与enu处理的C57BL/6J雄性杂交产生的10000多个G，后代的显性表型。迄今为止，我们已经获得了136个遗传证实的突变体，这些突变体表现出行为和形态缺陷。一些独立的突变系表现出相似的表型，表明它们可能代表相同基因的等位基因或相同遗传途径中的突变。它们对于揭示某些表型的分子机制具有巨大的潜力。
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 G, 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.