A regulatory toolbox of MiniPromoters to drive selective expression in the brain
A regulatory toolbox of MiniPromoters to drive selective expression in the brain
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DOI：
10.1073/pnas.1009158107
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发表时间：
2010-09-21
影响因子：
11.1
通讯作者：
Simpson, Elizabeth M.
中科院分区：
文献类型：
--
作者：
Portales-Casamar, Elodie;Swanson, Douglas J.;Simpson, Elizabeth M.
Pleiades启动子项目将全基因组生物信息学与大规模敲入小鼠生产和表达模式的组织学检查相结合，开发minipromoter和相关工具，旨在通过定向基因表达来研究和治疗大脑。对具有感兴趣的大脑表达模式的基因进行生物信息学分析，以描绘候选调控区域，然后将其纳入紧凑的人类微型启动子面板，以驱动对感兴趣的大脑区域和细胞类型的表达。利用胚胎干细胞中的单拷贝、同源重组“敲入蛋白”，每个MiniPromoter报告基因都与小鼠基因组中Hprt位点的5′位点直接整合。MiniPromoter的表达谱在转基因细胞或转基因小鼠生产后的小鼠大脑的分化试验中被表征。对成人大脑、眼睛和脊髓进行报告基因活性的组织学检查，并结合细胞类型特异性标记物进行染色以确定表达。公开的Pleiades MiniPromoter Project是促进大脑发育和治疗研究的关键资源。
The Pleiades Promoter Project integrates genomewide bioinformatics with large-scale knockin mouse production and histological examination of expression patterns to develop MiniPromoters and related tools designed to study and treat the brain by directed gene expression. Genes with brain expression patterns of interest are subjected to bioinformatic analysis to delineate candidate regulatory regions, which are then incorporated into a panel of compact human MiniPromoters to drive expression to brain regions and cell types of interest. Using single-copy, homologous-recombination "knockins" in embryonic stem cells, each MiniPromoter reporter is integrated immediately 5 ' of the Hprt locus in the mouse genome. MiniPromoter expression profiles are characterized in differentiation assays of the transgenic cells or in mouse brains following transgenic mouse production. Histological examination of adult brains, eyes, and spinal cords for reporter gene activity is coupled to costaining with cell-type-specific markers to define expression. The publicly available Pleiades MiniPromoter Project is a key resource to facilitate research on brain development and therapies.