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Non-canonical open reading frames encode functional proteins essential for cancer cell survival

biorxiv. 2020; 
John R. Prensner, Oana M. Enache,  Victor Luria,  Karsten Krug,  Karl R. Clauser,  Joshua M. Dempster,  Amir Karger,  Li Wang,  Karolina Stumbraite,  Vickie M. Wang,  Ginevra Botta,  Nicholas J. Lyons,  Amy Goodale,  Zohra Kalani,  Briana Fritchman,  Adam Brown,  Douglas Alan,  Thomas Green,  Xiaoping Yang,  Jacob D. Jaffe,  Jennifer A. Roth,  Federica Piccioni,  Marc W. Kirschner,  Zhe Ji,  David E. Root,  Todd R. Golub
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Molecular Biology Reagents The ORFeome library was then generated via insert synthesis and cloning of unique plasmid inserts consisting unique barcodes (Supplementary Table 22) by a commercial vendor (GenScript, Piscataway, NJ) in arrayed barcoded tube format. Get A Quote

摘要

A key question in genome research is whether biologically active proteins are restricted to the ∼20,000 canonical, well-annotated genes, or rather extend to the many non-canonical open reading frames (ORFs) predicted by genomic analyses. To address this, we experimentally interrogated 553 ORFs nominated in ribosome profiling datasets. Of these 553 ORFs, 57 (10%) induced a viability defect when the endogenous ORF was knocked out using CRISPR/Cas9 in 8 human cancer cell lines, 257 (46%) showed evidence of protein translation when ectopically expressed in HEK293T cells, and 401 (73%) induced gene expression changes measured by transcriptional profiling following ectopic expression across 4 cell types. CRISPR til... More

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