A CRISPR-based screen for Hedgehog signaling provides insights into ciliary function and ciliopathies.
A CRISPR-based screen for Hedgehog signaling provides insights into ciliary function and ciliopathies.
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DOI：
10.1038/s41588-018-0054-7
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发表时间：
2018-03
期刊：
影响因子：
30.8
通讯作者：
Nachury MV
中科院分区：
文献类型：
--
作者：
Breslow DK;Hoogendoorn S;Kopp AR;Morgens DW;Vu BK;Kennedy MC;Han K;Li A;Hess GT;Bassik MC;Chen JK;Nachury MV
初级纤毛组织刺猬信号和形成胚胎发育，其失调是纤毛疾病的统一原因。我们对Hedgehog信号进行了功能基因组筛选，方法是设计基于抗生素的Hedgehog反应细胞选择，并应用全基因组CRISPR介导的基因破坏。该屏幕可靠地识别纤毛信号所需的因素，几乎没有假阳性或假阴性。对HIT基因的研究揭示了几种纤毛结构的新成分，包括一种含有中心粒维持所需的δ-和ε-微管蛋白的蛋白质复合体。该屏幕还提供了一个公正的工具来对纤毛疾病进行分类，并揭示了许多先天性心脏病是由纤毛信号的丧失引起的。总体而言，我们的研究使系统分析睫毛功能和纤毛病变成为可能，并定义了一个通用的平台，用于通过基于CRISPR的筛查来解剖信号通路。
The primary cilium organizes Hedgehog signaling and shapes embryonic development, and its dysregulation is the unifying cause of ciliopathies. We conducted a functional genomic screen for Hedgehog signaling by engineering antibiotic-based selection of Hedgehog-responsive cells and applying genome-wide CRISPR-mediated gene disruption. The screen robustly identifies factors required for ciliary signaling with few false positives or false negatives. Characterization of hit genes uncovers novel components of several ciliary structures, including a protein complex containing δ- and ε-tubulin that is required for centriole maintenance. The screen also provides an unbiased tool for classifying ciliopathies and reveals that many congenital heart disorders are caused by loss of ciliary signaling. Collectively, our study enables a systematic analysis of ciliary function and of ciliopathies and also defines a versatile platform for dissecting signaling pathways through CRISPR-based screening.
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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