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bioRxiv · 10.1101/171819

C-BERST: Defining subnuclear proteomic landscapes at genomic elements with dCas9-APEX2

Gao, X. D.·Tu, L.-C.·Mir, A.·Dekker, J.·Shaffer, S. A.·Zhu, L. J.·Wolfe, S. A.·Sontheimer, E. J.
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Abstract

Mapping proteomic composition at distinct genomic loci and subnuclear landmarks in living cells has been a long-standing challenge. Here we report that dCas9-APEX2 Biotinylation at genomic Elements by Restricted Spatial Tagging (C-BERST) allows the rapid, unbiased mapping of proteomes near defined genomic loci, as demonstrated for telomeres and centromeres. By combining the spatially restricted enzymatic tagging enabled by APEX2 with programmable DNA targeting by dCas9, C-BERST has successfully identified nearly 50% of known telomere-associated factors and many known centromere-associated factors. We also identified and validated SLX4IP and RPA3 as telomeric factors, confirming C-BERSTs utility as a discovery platform. C-BERST enables the rapid, high-throughput identification of proteins associated with specific sequences, facilitating annotation of these factors and their roles in nuclear and chromosome biology.

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Gao, X. D., Tu, L.-C., Mir, A., Dekker, J., Shaffer, S. A., Zhu, L. J., Wolfe, S. A., Sontheimer, E. J.. 2017-08-02. C-BERST: Defining subnuclear proteomic landscapes at genomic elements with dCas9-APEX2. https://doi.org/10.1101/171819

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