Abstract
Identification of genes regulating fat accumulation is important for basic and medical research; genetic screening for those genes in Caenorhabditis elegans, a widely used model organism, requires in vivo quantification of lipids. We demonstrated RNA interference screening based on quantitative imaging of lipids with label-free stimulated Raman scattering (SRS) microscopy, which overcomes major limitations of coherent anti-Stokes Raman scattering microscopy. Our screening yielded eight new genetic regulators of fat storage.
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Acknowledgements
We thank members of the H. Hutter laboratory (Simon Fraser University) for providing the nre-1(hd20)lin-15b(hd126) strain, members of the C. Sztalryd lab (University of Maryland School of Medicine) for providing perilipin A, ADFP and LSDP5 plasmids, J. Melo and Y. Hao for helping with the membrane receptor RNAi library, A. Soukas and H. Mak for helping with the nuclear hormone receptor RNAi library, Z. Shi for helping with RNAi clone sequencing, X. Zhang and B. Saar for helping on imaging instrumentation, P. Iakova and A. Folick for helping with mammalian cell transfection and C. He and J. Kang for gas chromatography analysis. This work was supported by US National Institutes of Health grants AG034988 (M.C.W.) and EB010244 (X.S.X.) and a predoctoral fellowship from Boehringer Ingelheim Fonds (C.W.F.).
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M.C.W., W.M. and X.S.X. conceived the study; M.C.W. and W.M. designed the experiments; M.C.W., W.M. and C.W.F. performed the experiments; M.C.W. analyzed the data; M.C.W., W.M., G.R. and X.S.X. wrote the manuscript.
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Wang, M., Min, W., Freudiger, C. et al. RNAi screening for fat regulatory genes with SRS microscopy. Nat Methods 8, 135–138 (2011). https://doi.org/10.1038/nmeth.1556
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DOI: https://doi.org/10.1038/nmeth.1556
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