Massively parallel high-order combinatorial genetics in human cells
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Author(s)
Wong, Siu Lun
Choi, Ching Gee
Cheng, Allen
Purcell, Oliver
Lu, Timothy K
Date Issued
August 2015
Journal
Nature Biotechnology
Publisher
Nature Publishing Group
Citation
Wong, Alan S L et al. “Massively Parallel High-Order Combinatorial Genetics in Human Cells.” Nature Biotechnology 33, 9 (August 2015): 952–961 © 2015 Nature America, Inc
Version
Author's final manuscript
Abstract
The systematic functional analysis of combinatorial genetics has been limited by the throughput that can be achieved and the order of complexity that can be studied. To enable massively parallel characterization of genetic combinations in human cells, we developed a technology for rapid, scalable assembly of high-order barcoded combinatorial genetic libraries that can be quantified with high-throughput sequencing. We applied this technology, combinatorial genetics en masse (CombiGEM), to create high-coverage libraries of 1,521 two-wise and 51,770 three-wise barcoded combinations of 39 human microRNA (miRNA) precursors. We identified miRNA combinations that synergistically sensitize drug-resistant cancer cells to chemotherapy and/or inhibit cancer cell proliferation, providing insights into complex miRNA networks. More broadly, our method will enable high-throughput profiling of multifactorial genetic combinations that regulate phenotypes of relevance to biomedicine, biotechnology and basic science.
MIT Department
Massachusetts Institute of Technology. Synthetic Biology Center
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Massachusetts Institute of Technology. Research Laboratory of Electronics
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/nbt.3326
