A Modular Assembly Platform for Rapid Generation of DNA Constructs
Name
Akama-Garren-2016-Modular Assembly.pdf
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1.4 MB
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Author(s)
Tammela, Tuomas
Chang, Gregory P.
Wagner, Bethany L.
Lee, Da-Yae
Papagiannakopoulos, Thales
Xue, Wen
Joshi, Nik
Akama-Garren, Elliot
Rideout, William M.
Jacks, Tyler E
Date Issued
February 2016
Journal
Scientific Reports
Publisher
Nature Publishing Group
Citation
Akama-Garren, Elliot H., Nikhil S. Joshi, Tuomas Tammela, Gregory P. Chang, Bethany L. Wagner, Da-Yae Lee, William M. Rideout III, Thales Papagiannakopoulos, Wen Xue, and Tyler Jacks. “A Modular Assembly Platform for Rapid Generation of DNA Constructs.” Scientific Reports 6 (February 18, 2016): 16836.
Version
Final published version
Abstract
Traditional cloning methods have limitations on the number of DNA fragments that can be simultaneously manipulated, which dramatically slows the pace of molecular assembly. Here we describe GMAP, a Gibson assembly-based modular assembly platform consisting of a collection of promoters and genes, which allows for one-step production of DNA constructs. GMAP facilitates rapid assembly of expression and viral constructs using modular genetic components, as well as increasingly complicated genetic tools using contextually relevant genomic elements. Our data demonstrate the applicability of GMAP toward the validation of synthetic promoters, identification of potent RNAi constructs, establishment of inducible lentiviral systems, tumor initiation in genetically engineered mouse models, and gene-targeting for the generation of knock-in mice. GMAP represents a recombinant DNA technology designed for widespread circulation and easy adaptation for other uses, such as synthetic biology, genetic screens, and CRISPR-Cas9.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1038/srep16836
