Inverse drug screens: A rapid and inexpensive method for implicating molecular targets
Inverse drug screens: A rapid and inexpensive method for implicating molecular targets
复制标题
DOI：
10.1002/dvg.20246
复制
发表时间：
2006-11-01
期刊：
影响因子：
1.5
通讯作者：
Levin, Michael
中科院分区：
文献类型：
--
作者：
Adams, Dany S.;Levin, Michael
鉴定在某些特定过程中起作用的基因产物是发育生物学的共同目标。虽然使用药物化合物来探测生物系统在畸形学和毒理学中具有非常长的历史，但系统的分层药物筛选尚未被发育生物学界利用。这种“化学遗传学”方法可以极大地有利于胚胎和再生系统的研究，我们已经正式制定了一项战略，使用已知的药理学化合物牵连特定的分子候选人在任何选择的生物现象。利用可以在许多领域（如离子转运、神经递质功能、代谢和细胞骨架）中施加于药物试剂的分层结构，任何测定都可以作为二分搜索算法进行。这种反向药物筛选方法比大量药物的详尽测试更有效，并且揭示了可管理数量的特定分子候选物的身份，然后可以使用更昂贵和特定的分子试剂进行验证和靶向。在这里，我们描述的过程中，这种功能丧失的屏幕，并说明其使用在发现新的生物电和胚胎图案的神经元能机制。该技术是对现有分子筛选策略的廉价且快速的补充。此外，它适用于母体蛋白质，以及传统遗传筛选不可行的模式物种，显著延长了识别生物过程中关键内源性参与者的机会。
Identification of gene products that function in some specific process of interest is a common goal in developmental biology. Although use of drug compounds to probe biological systems has a very long history in teratology and toxicology, systematic hierarchical drug screening has not been capitalized upon by the developmental biology community. This "chemical genetics" approach can greatly benefit the study of embryonic and regenerative systems, and we have formalized a strategy for using known pharmacological compounds to implicate specific molecular candidates in any chosen biological phenomenon. Taking advantage of a hierarchical structure that can be imposed on drug reagents in a number of fields such as ion transport, neurotransmitter function, metabolism, and cytoskeleton, any assay can be carried out as a binary search algorithm. This inverse drug screen methodology is much more efficient than exhaustive testing of large numbers of drugs, and reveals the identity of a manageable number of specific molecular candidates that can then be validated and targeted using more expensive and specific molecular reagents. Here, we describe the process of this loss-of-function screen and illustrate its use in uncovering novel bioelectrical and serotonergic mechanisms in embryonic patterning. This technique is an inexpensive and rapid complement to existing molecular screening strategies. Moreover, it is applicable to maternal proteins, and model species in which traditional genetic screens are not feasible, significantly extending the opportunities to identify key endogenous players in biological processes.