Abstract
We present a FISH-based method for detecting single-nucleotide variants (SNVs) in exons and introns on individual RNA transcripts with high efficiency. We used this method to quantify allelic expression in cell populations and in single cells, and also to distinguish maternal from paternal chromosomes in single cells.
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Change history
14 August 2013
In the version of this article initially published online, a bar in panel d of Figure 1 was left blank. It should have been filled with the appropriate colors to indicate the number of transcripts derived from wild-type (blue) or mutant (orange) RNA and the number of unclassified transcripts (colorless). The error has been corrected for the print, PDF and HTML versions of the article.
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Acknowledgements
We thank Biosearch technologies for providing many of the reagents used in our assays and G. Nair for many discussions about statistics. We acknowledge the US National Institutes of Health Director's New Innovator Award (1DP2OD008514; M.J.L., P.G., Y.W. and A.R.) and a Burroughs-Wellcome Fund Career Award at the Scientific Interface (A.R.) for supporting our work.
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M.J.L. conceived of the method with guidance from A.R. M.J.L. performed the image analysis and P.G. performed the statistical analysis. M.J.L., Y.W. and P.G. performed the experiments. M.J.L., P.G. and A.R. wrote the paper.
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A.R. and M.J.L. are inventors on a US provisional patent 61/785,498 on the method described in this paper, and receive royalties related to this and other related patents.
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Supplementary Figures 1–8, Supplementary Tables 1 and 2, and Supplementary Note 1 (PDF 1655 kb)
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Levesque, M., Ginart, P., Wei, Y. et al. Visualizing SNVs to quantify allele-specific expression in single cells. Nat Methods 10, 865–867 (2013). https://doi.org/10.1038/nmeth.2589
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DOI: https://doi.org/10.1038/nmeth.2589
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