Journal article
Model-Driven Engineering of Gene Expression from RNA Replicons
ACS synthetic biology, Vol.4(1), pp.48-56
01/16/2015
DOI: 10.1021/sb500173f
PMID: 24877739
Abstract
RNA replicons are an emerging platform for engineering synthetic biological systems. Replicons self-amplify, can provide persistent high-level expression of proteins even from a small initial dose, and, unlike DNA vectors, pose minimal risk of chromosomal integration. However, no quantitative model sufficient for engineering levels of protein expression from such replicon systems currently exists. Here, we aim to enable the engineering of multigene expression from more than one species of replicon by creating a computational model based on our experimental observations of the expression dynamics in single- and multireplicon systems. To this end, we studied fluorescent protein expression in baby hamster kidney (BHK-21) cells using a replicon derived from Sindbis virus (SINV). We characterized expression dynamics for this platform based on the doseresponse of a single species of replicon over 50 h and on a titration of two cotransfected replicons expressing different fluorescent proteins. From this data, we derive a quantitative model of multireplicon expression and validate it by designing a variety of three-replicon systems, with profiles that match desired expression levels. We achieved a mean error of 1.7-fold on a 1000-fold range, thus demonstrating how our model can be applied to precisely control expression levels of each Sindbis replicon species in a system.
Details
- Title: Subtitle
- Model-Driven Engineering of Gene Expression from RNA Replicons
- Creators
- Jacob Beal - RTXTyler E. Wagner - Boston UniversityTasuku Kitada - Massachusetts Institute of TechnologyOdisse Azizgolshani - University of California, Los AngelesJordan Moberg Parker - University of California, Los AngelesDouglas Densmore - Boston UniversityRon Weiss - Massachusetts Institute of Technology
- Resource Type
- Journal article
- Publication Details
- ACS synthetic biology, Vol.4(1), pp.48-56
- Publisher
- Amer Chemical Soc
- DOI
- 10.1021/sb500173f
- PMID
- 24877739
- ISSN
- 2161-5063
- eISSN
- 2161-5063
- Number of pages
- 9
- Grant note
- W911NF-11-054 / DARPA; United States Department of Defense; Defense Advanced Research Projects Agency (DARPA)
- Language
- English
- Date published
- 01/16/2015
- Academic Unit
- Electrical and Computer Engineering
- Record Identifier
- 9984627187702771
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