General approach for in vivo recovery of cell type-specific effector gene sets.

General approach for in vivo recovery of cell type-specific effector gene sets.
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DOI:
10.1101/gr.167668.113
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发表时间:
2014-05
期刊:
影响因子:
7
通讯作者:
Davidson EH
Davidson EH
中科院分区:
生物学1区
文献类型:
--
作者:
Barsi JC;Tu Q;Davidson EH

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差异表达的细胞类型特异性效应基因集是解决生物学中多个重要问题的关键,从发育基因调控网络(GRN)的理论方面到各种实际应用。尽管已通过各种方法回收并分析了感兴趣的单个细胞类型,但从整个发育生物体中系统回收多种细胞类型特异性基因集仍然存在问题。在这里,我们描述了一种使用海胆胚胎的通用方法,海胆胚胎是一种选择材料,因为该模型系统已经解决了大规模的 GRN。该方法利用胚胎给定细胞表达的调节状态来定义细胞类型,并包括荧光激活细胞分选 (FACS) 程序,该程序不会导致转录物表示的干扰。我们通过对分离的胚胎成骨细胞中表达的转录组进行空间和定性分析来广泛验证该方法,并因此生成了原型细胞类型特异性转录组数据库。
Differentially expressed, cell type-specific effector gene sets hold the key to multiple important problems in biology, from theoretical aspects of developmental gene regulatory networks (GRNs) to various practical applications. Although individual cell types of interest have been recovered by various methods and analyzed, systematic recovery of multiple cell type-specific gene sets from whole developing organisms has remained problematic. Here we describe a general methodology using the sea urchin embryo, a material of choice because of the large-scale GRNs already solved for this model system. This method utilizes the regulatory states expressed by given cells of the embryo to define cell type and includes a fluorescence activated cell sorting (FACS) procedure that results in no perturbation of transcript representation. We have extensively validated the method by spatial and qualitative analyses of the transcriptome expressed in isolated embryonic skeletogenic cells and as a consequence, generated a prototypical cell type-specific transcriptome database.
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