原创力文档a high-throughput assay for sirna-based circadian screens in human u2os cellssirna-based昼夜屏幕的高通量分析人类u2os细胞.pdfVIP
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a high-throughput assay for sirna-based circadian screens in human u2os cellssirna-based昼夜屏幕的高通量分析人类u2os细胞
A High-Throughput Assay for siRNA-Based Circadian
Screens in Human U2OS Cells
1,2. 1 1.
Christopher Vollmers , Satchidananda Panda , Luciano DiTacchio *
1 Regulatory Biology Laboratory, Salk Institute for Biological Studies, La Jolla, California, United States of America, 2 Heidelberg University Biochemistry Center (BZH),
Heidelberg, Germany
Abstract
The advent of siRNA-based screens has revolutionized the efficiency by which functional components of biological
processes are identified. A notable exception has been the field of mammalian circadian rhythms. Here, we outline a
medium- to high-throughput siRNA-based approach that, in combination with real-time bioluminescence measurement of
a circadian reporter gene, can be utilized to elucidate the effects of gene knockdown across several days in human cells.
Citation: Vollmers C, Panda S, DiTacchio L (2008) A High-Throughput Assay for siRNA-Based Circadian Screens in Human U2OS Cells. PLoS ONE 3(10): e3457.
doi:10.1371/journal.pone.0003457
Editor: Michael N. Nitabach, Yale School of Medicine, United States of America
Received August 25, 2008; Accepted September 23, 2008; Published October 20, 2008
Copyright: 2008 Vollmers et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by the Pew Scholars Program in Biomedical Sciences, Dana Foundation, Whitehall Foundation and NIH grants EY016807 (to
S.P.), and by NRSA 1F32GM082083-01 (to L.D.)
Competing Interests: The authors have declared that no competing int
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