The Tribolium castaneum cell line TcA: a new tool kit for cell biology.
The Tribolium castaneum cell line TcA: a new tool kit for cell biology.
复制标题
Tribolium Castaneum细胞系TCA：一种用于细胞生物学的新工具套件。
DOI：
10.1038/srep06840
复制
发表时间：
2014-10-30
影响因子：
4.6
通讯作者：
Park Y
中科院分区：
文献类型：
--
作者：
Silver K;Jiang H;Fu J;Phillips TW;Beeman RW;Park Y
赤拟谷盗（Tribolium castaneum）是一种重要的农业害虫，已被广泛用作模式生物。最近，从红粉甲虫的晚期蛹中开发了贴壁细胞系（BCIRL-TcA-CLG 1或TcA）。TcA细胞的下一代转录组测序证明了与几丁质代谢、免疫应答以及细胞和系统RNAi途径中的专门功能相关的多种基因的表达。因此，我们评估了TcA细胞对dsRNA启动RNAi应答的敏感性。TcA细胞对微量dsRNA高度敏感，最小有效剂量为100 pg/mL，导致基因表达的显著抑制。 我们还开发了含有两个TcA特异性启动子的质粒，来自40 S核糖体蛋白亚基的启动子（TC 006550）和来自热休克蛋白68 a和B之间的基因间隔的双向热休克启动子（TcHS 70）。这些启动子已用于提供报告蛋白的高水平的组成型（TC 006550）或诱导型（TcHS 70）基因表达。我们的研究结果表明，TcA细胞系，其对RNAi和功能TcA特异性启动子的敏感性，是研究基本分子和生理问题的宝贵资源。
The red flour beetle, Tribolium castaneum, is an agriculturally important insect pest that has been widely used as a model organism. Recently, an adherent cell line (BCIRL-TcA-CLG1 or TcA) was developed from late pupae of the red flour beetle. Next generation transcriptome sequencing of TcA cells demonstrated expression of a wide variety of genes associated with specialized functions in chitin metabolism, immune responses and cellular and systemic RNAi pathways. Accordingly, we evaluated the sensitivity of TcA cells to dsRNA to initiate an RNAi response. TcA cells were highly sensitive to minute amounts of dsRNA, with a minimum effective dose of 100 pg/mL resulting in significant suppression of gene expression. We have also developed a plasmid containing two TcA-specific promoters, the promoter from the 40S ribosomal protein subunit (TC006550) and a bi-directional heat shock promoter (TcHS70) from the intergenic space between heat shock proteins 68a and b. These promoters have been employed to provide high levels of either constitutive (TC006550) or inducible (TcHS70) gene expression of the reporter proteins. Our results show that the TcA cell line, with its sensitivity to RNAi and functional TcA-specific promoters, is an invaluable resource for studying basic molecular and physiological questions.
登录
查看更多内容
影响因子：
12.3
作者：
Tomoyasu Y;Miller SC;Tomita S;Schoppmeier M;Grossmann D;Bucher G
通讯作者：
Bucher G
影响因子：
4.6
作者：
Kayukawa, Takumi;Tateishi, Ken;Shinoda, Tetsuro
通讯作者：
Shinoda, Tetsuro
影响因子：
4.5
作者：
Shih, Joseph D.;Fitzgerald, Michael C.;Hunter, Craig P.
通讯作者：
Hunter, Craig P.
影响因子：
14.9
作者：
Kim HS;Murphy T;Xia J;Caragea D;Park Y;Beeman RW;Lorenzen MD;Butcher S;Manak JR;Brown SJ
通讯作者：
Brown SJ
影响因子：
64.8
作者：
Richards, Stephen;Gibbs, Richard A.;Grossmann, Daniela
通讯作者：
Grossmann, Daniela