FRET and BRET-based biosensors in live cell compound screens.

FRET and BRET-based biosensors in live cell compound screens.
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活细胞化合物筛选中基于 FRET 和 BRET 的生物传感器。

DOI:
10.1007/978-1-62703-622-1_17
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发表时间:
2014
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Zhang,Jin
Zhang,Jin
中科院分区:
--
文献类型:
--
作者:
Robinson,KatieHerbst;Yang,JessicaR;Zhang,Jin

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活细胞化合物筛选与遗传编码的荧光或生物发光为基础的生物传感器提供了一个潜在的强大的方法,以确定新的监管机构的信号事件的利益。特别地,活细胞中的化合物筛选具有额外的益处,即整个信号传导网络保持完整,因此筛选不仅针对单个感兴趣的分子,而且针对信号传导网络内的任何分子,其可以调节使用中的生物传感器报告的不同信号传导事件。此外,只有细胞可渗透的或作用于细胞表面受体的分子将被鉴定为“命中”,因此减少了化合物在细胞渗透方面的进一步优化。在这里,我们讨论了一个详细的协议,用于使用基因编码的生物传感器在活细胞中的96孔格式的高通量化合物的屏幕和识别的小分子,调节感兴趣的信号事件的执行。
Live cell compound screening with genetically encoded fluorescence or bioluminescence-based biosensors offers a potentially powerful approach to identify novel regulators of a signaling event of interest. In particular, compound screening in living cells has the added benefit that the entire signaling network remains intact, and thus the screen is not just against a single molecule of interest but against any molecule within the signaling network that may modulate the distinct signaling event reported by the biosensor in use. Furthermore, only molecules that are cell permeable or act at cell surface receptors will be identified as “hits,” thus reducing further optimization of the compound in terms of cell penetration. Here we discuss a detailed protocol for using genetically encoded biosensors in living cells in a 96-well format for the execution of high throughput compound screens and the identification of small molecules which modulate a signaling event of interest.
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发表时间: 2006-01-01
影响因子: 4
作者:
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发表时间: 2017
期刊: --
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