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A High-Throughput Microfluidic Platform for Mammalian Cell Transfection and Culturing.
- Source :
-
Scientific reports [Sci Rep] 2016 Mar 31; Vol. 6, pp. 23937. Date of Electronic Publication: 2016 Mar 31. - Publication Year :
- 2016
-
Abstract
- Mammalian synthetic biology could be augmented through the development of high-throughput microfluidic systems that integrate cellular transfection, culturing, and imaging. We created a microfluidic chip that cultures cells and implements 280 independent transfections at up to 99% efficiency. The chip can perform co-transfections, in which the number of cells expressing each protein and the average protein expression level can be precisely tuned as a function of input DNA concentration and synthetic gene circuits can be optimized on chip. We co-transfected four plasmids to test a histidine kinase signaling pathway and mapped the dose dependence of this network on the level of one of its constituents. The chip is readily integrated with high-content imaging, enabling the evaluation of cellular behavior and protein expression dynamics over time. These features make the transfection chip applicable to high-throughput mammalian protein and synthetic biology studies.
- Subjects :
- Cell Culture Techniques instrumentation
Gene Expression Regulation
HEK293 Cells
Histidine Kinase genetics
Histidine Kinase metabolism
Humans
Microfluidics instrumentation
Optical Imaging instrumentation
Plasmids chemistry
Signal Transduction
Transfection
Cell Culture Techniques methods
Lab-On-A-Chip Devices
Microfluidics methods
Optical Imaging methods
Plasmids metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 6
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 27030663
- Full Text :
- https://doi.org/10.1038/srep23937