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A public genome-scale lentiviral expression library of human ORFs


Functional characterization of the human genome requires tools for systematically modulating gene expression in both loss-of-function and gain-of-function experiments. We describe the production of a sequence-confirmed, clonal collection of over 16,100 human open-reading frames (ORFs) encoded in a versatile Gateway vector system. Using this ORFeome resource, we created a genome-scale expression collection in a lentiviral vector, thereby enabling both targeted experiments and high-throughput screens in diverse cell types.

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Figure 1: Overview of hORFeome V8.1.
Figure 2: Performance of the CCSB-Broad lentiviral expression library.

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We thank B. Piqani, I. Budianto, D. Szeto, T. Hirozane-Kishikawa, V. Swearingen, A. MacWilliams, T. Nieland, S. Hoang, J. Bochicchio, S. Young, A. Berlin, C. Russ, M. Garber and members of the Broad Institute Genetic Sequencing Platform who provided technical assistance or advice throughout this project, and J. Zhao, T. Roberts and T. Golub for participation in kinase ORFs subcollection generation. This work was supported by Broad Institute Scientific Planning and Allocation of Resources Committee funding, The Ellison Foundation (D.E.H. and M.V.), Dana-Farber Cancer Institute sponsored research funds to CCSB and Center for Cancer Genome Discovery and US National Institutes of Health R33 CA128625 (W.C.H., D.E.R. and D.E.H.).

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Authors and Affiliations



J.S.B., Xia. Y. and D.E.R. wrote the manuscript and, together with D.E.H., K.S.-A. and M.V., designed and supervised the process of creating clonal sequenced ORF collections. Xin. Y., D.B., L.G., J.S.B., S.R.T., H.H., R.R.M., K.S.-A. and C.M.J. generated the starting collections of ORFs. R.L., O.A., J.S.B., C.N., Xia. Y., S.J.S., S.R.T. and C.M.J. created the final libraries, DNA, viruses and pLX vectors, and evaluated expression. T.H., Y.S., C.F., C.L., J.S.B., T.B., T.M.G., Xia. Y. and D.E.R. performed bioinformatic analyses. M.V., W.C.H., D.E.H. and D.E.R. supervised the project.

Corresponding authors

Correspondence to Marc Vidal, William C Hahn, David E Hill or David E Root.

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The authors declare no competing financial interests.

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Supplementary Figures 1–12, Supplementary Tables 1–4; Supplementary Notes 1–8 (PDF 2703 kb)

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Yang, X., Boehm, J., Yang, X. et al. A public genome-scale lentiviral expression library of human ORFs. Nat Methods 8, 659–661 (2011).

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