Development of an efficient screening system to identify novel bone metabolism-related genes using the exchangeable gene trap mutagenesis mouse models.
Development of an efficient screening system to identify novel bone metabolism-related genes using the exchangeable gene trap mutagenesis mouse models.
复制标题
开发有效的筛查系统，使用可交换基因陷阱诱变小鼠模型鉴定与骨代谢相关的新基因。
DOI：
10.1038/srep40692
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发表时间：
2017-01-20
影响因子：
4.6
通讯作者：
Chosa E
中科院分区：
文献类型：
--
作者：
Kurogi S;Sekimoto T;Funamoto T;Ota T;Nakamura S;Nagai T;Nakahara M;Yoshinobu K;Araki K;Araki M;Chosa E
尽管有许多关于骨代谢的遗传学研究，但缺乏对具体机制的理解。我们开发了一个有效的筛选系统，以确定新的基因参与骨代谢的突变小鼠品系登记的可交换基因陷阱克隆（EGTC）数据库。从EGTC数据库中的1278个诱捕克隆中，根据“EST profile”、“X-gal”、“Related article”和“Novel gene”确定了52个候选株系。对于第二次筛选，对候选品系进行骨形态测量分析、生物力学强度分析、骨X-gal染色等。42条雄性诱捕线（80.8%）显示骨形态测量分析或生物力学强度分析异常。在筛选过程中，X-gal染色是显著有效的（P = 0.0057）。例如，使用筛选系统选择的Lbr和Nedd 4陷阱系显示出显著的骨减少和脆性，表明与成骨细胞分化的关系。这种使用EGTC小鼠系的筛选系统对于鉴定参与骨代谢的新基因是非常有效的。使用这种筛选方法鉴定为异常的基因捕获系极有可能捕获骨代谢的重要基因。这些筛选出的诱捕鼠将作为骨研究的新生物资源。
Despite numerous genetic studies on bone metabolism, understanding of the specific mechanisms is lacking. We developed an efficient screening system to identify novel genes involved in bone metabolism using mutant mouse strains registered with the Exchangeable Gene Trap Clones (EGTC) database. From 1278 trap clones in the EGTC database, 52 candidate lines were selected in the first screening, determined based on “EST profile”, “X-gal”, “Related article”, and “Novel gene”. For the second screening, bone morphometric analysis, biomechanical strength analysis, bone X-gal staining, etc. were performed on candidate lines. Forty-two male trap lines (80.8%) showed abnormalities with either bone morphometric analysis or biomechanical strength analysis. In the screening process, X-gal staining was significantly efficient (P = 0.0057). As examples, Lbr and Nedd4 trap lines selected using the screening system showed significant bone decrease and fragility, suggesting a relationship with osteoblast differentiation. This screening system using EGTC mouse lines is extremely efficient for identifying novel genes involved in bone metabolism. The gene trap lines identified as abnormal using this screening approach are highly likely to trap important genes for bone metabolism. These selected trap mice will be valuable for use as novel bio-resources in bone research.