Rapid, modular and reliable construction of complex mammalian gene circuits
We developed a framework for quick and reliable construction of complex gene circuits for genetically engineering mammalian cells. Our hierarchical framework is based on a novel nucleotide addressing system for defining the position of each part in an overall circuit. With this framework, we demonst...
| Main Authors: | , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Oxford University Press
2013
|
| Online Access: | http://hdl.handle.net/1721.1/80718 https://orcid.org/0000-0003-0396-2443 |
| _version_ | 1826194377256468480 |
|---|---|
| author | Guye, Patrick Li, Yinqing Wroblewska, Liliana Duportet, Xavier Weiss, Ron |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Guye, Patrick Li, Yinqing Wroblewska, Liliana Duportet, Xavier Weiss, Ron |
| author_sort | Guye, Patrick |
| collection | MIT |
| description | We developed a framework for quick and reliable construction of complex gene circuits for genetically engineering mammalian cells. Our hierarchical framework is based on a novel nucleotide addressing system for defining the position of each part in an overall circuit. With this framework, we demonstrate construction of synthetic gene circuits of up to 64 kb in size comprising 11 transcription units and 33 basic parts. We show robust gene expression control of multiple transcription units by small molecule inducers in human cells with transient transfection and stable chromosomal integration of these circuits. This framework enables development of complex gene circuits for engineering mammalian cells with unprecedented speed, reliability and scalability and should have broad applicability in a variety of areas including mammalian cell fermentation, cell fate reprogramming and cell-based assays. |
| first_indexed | 2024-09-23T09:55:03Z |
| format | Article |
| id | mit-1721.1/80718 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:55:03Z |
| publishDate | 2013 |
| publisher | Oxford University Press |
| record_format | dspace |
| spelling | mit-1721.1/807182022-09-30T17:40:37Z Rapid, modular and reliable construction of complex mammalian gene circuits Guye, Patrick Li, Yinqing Wroblewska, Liliana Duportet, Xavier Weiss, Ron Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Guye, Patrick Li, Yinqing Wroblewska, Liliana Duportet, Xavier Weiss, Ron We developed a framework for quick and reliable construction of complex gene circuits for genetically engineering mammalian cells. Our hierarchical framework is based on a novel nucleotide addressing system for defining the position of each part in an overall circuit. With this framework, we demonstrate construction of synthetic gene circuits of up to 64 kb in size comprising 11 transcription units and 33 basic parts. We show robust gene expression control of multiple transcription units by small molecule inducers in human cells with transient transfection and stable chromosomal integration of these circuits. This framework enables development of complex gene circuits for engineering mammalian cells with unprecedented speed, reliability and scalability and should have broad applicability in a variety of areas including mammalian cell fermentation, cell fate reprogramming and cell-based assays. Synthetic Biology Engineering Research Center (SA5284-11210) United States. Defense Advanced Research Projects Agency (HR0011-12-C-0067) United States. Defense Advanced Research Projects Agency (DARPA-BAA-11-23) National Science Foundation (U.S.) (CBET-0939511) National Institutes of Health (U.S.). (5-R01-CA155320-02) 2013-09-13T15:04:09Z 2013-09-13T15:04:09Z 2013-07 2013-06 Article http://purl.org/eprint/type/JournalArticle 0305-1048 1362-4962 http://hdl.handle.net/1721.1/80718 Guye, P., Y. Li, L. Wroblewska, X. Duportet, and R. Weiss. “Rapid, modular and reliable construction of complex mammalian gene circuits.” Nucleic Acids Research (July 11, 2013). https://orcid.org/0000-0003-0396-2443 en_US http://dx.doi.org/10.1093/nar/gkt605 Nucleic Acids Research Creative Commons Attribution 3.0 http://creativecommons.org/licenses/by/3.0/ application/pdf Oxford University Press Oxford |
| spellingShingle | Guye, Patrick Li, Yinqing Wroblewska, Liliana Duportet, Xavier Weiss, Ron Rapid, modular and reliable construction of complex mammalian gene circuits |
| title | Rapid, modular and reliable construction of complex mammalian gene circuits |
| title_full | Rapid, modular and reliable construction of complex mammalian gene circuits |
| title_fullStr | Rapid, modular and reliable construction of complex mammalian gene circuits |
| title_full_unstemmed | Rapid, modular and reliable construction of complex mammalian gene circuits |
| title_short | Rapid, modular and reliable construction of complex mammalian gene circuits |
| title_sort | rapid modular and reliable construction of complex mammalian gene circuits |
| url | http://hdl.handle.net/1721.1/80718 https://orcid.org/0000-0003-0396-2443 |
| work_keys_str_mv | AT guyepatrick rapidmodularandreliableconstructionofcomplexmammaliangenecircuits AT liyinqing rapidmodularandreliableconstructionofcomplexmammaliangenecircuits AT wroblewskaliliana rapidmodularandreliableconstructionofcomplexmammaliangenecircuits AT duportetxavier rapidmodularandreliableconstructionofcomplexmammaliangenecircuits AT weissron rapidmodularandreliableconstructionofcomplexmammaliangenecircuits |