A platform for rapid prototyping of synthetic gene networks in mammalian cells
Name
Duportet-2014-A platform for rapid.pdf
Size
3.74 MB
Format
Adobe PDF
Checksum (MD5)
a2d1faede87727a4d81c90405a6742a5
Author(s)
Wroblewska, Liliana
Duportet, Xavier
Guye, Patrick
Li, Yinqing
Weiss, Ron
Eyquem, Justin
Rieders, Julianne
Rimchala, Tharathorn
Batt, Gregory
Date Issued
November 2014
Journal
Nucleic Acids Research
Publisher
Oxford University Press
Citation
Duportet, Xavier, Liliana Wroblewska, Patrick Guye, Yingqing Li, Justin Eyquem, Julianne Rieders, Tharathorn Rimchala, Gregory Batt, and Ron Weiss. “A Platform for Rapid Prototyping of Synthetic Gene Networks in Mammalian Cells.” Nucleic Acids Research 42, no. 21 (November 5, 2014): 13440–13451.
Version
Final published version
Abstract
Mammalian synthetic biology may provide novel therapeutic strategies, help decipher new paths for drug discovery and facilitate synthesis of valuable molecules. Yet, our capacity to genetically program cells is currently hampered by the lack of efficient approaches to streamline the design, construction and screening of synthetic gene networks. To address this problem, here we present a framework for modular and combinatorial assembly of functional (multi)gene expression vectors and their efficient and specific targeted integration into a well-defined chromosomal context in mammalian cells. We demonstrate the potential of this framework by assembling and integrating different functional mammalian regulatory networks including the largest gene circuit built and chromosomally integrated to date (6 transcription units, 27kb) encoding an inducible memory device. Using a library of 18 different circuits as a proof of concept, we also demonstrate that our method enables one-pot/single-flask chromosomal integration and screening of circuit libraries. This rapid and powerful prototyping platform is well suited for comparative studies of genetic regulatory elements, genes and multi-gene circuits as well as facile development of libraries of isogenic engineered cell lines.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Massachusetts Institute of Technology. Synthetic Biology Center
Terms of Use
Creative Commons Attribution
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1093/nar/gku1082
