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Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells
Published on: January 7, 2020
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Massively parallel RNA device engineering in mammalian cells with RNA-Seq
Joy S Xiang1, Matias Kaplan1, Peter Dykstra1
1Department of Bioengineering, 443 Via Ortega, MC 4245, Stanford University, Stanford, CA, 94305, USA.
Nature Communications
|September 25, 2019
Summary
Researchers developed a high-throughput RNA-Seq assay to engineer synthetic RNA genetic devices in mammalian cells. This method identified novel ribozyme switches responsive to small molecules, enhancing genetic engineering tools.
Area of Science:
- Synthetic Biology
- Molecular Biology
- Genetics
Background:
- Synthetic RNA genetic devices offer dynamic gene regulation across cell types.
- Limited assay throughput in mammalian cells impedes rapid engineering of RNA devices.
Purpose of the Study:
- To develop a quantitative, rapid, high-throughput mammalian cell-based RNA-Seq assay for engineering RNA devices.
- To identify novel small molecule-responsive ribozyme-based RNA devices.
Main Methods:
- Developed a high-throughput RNA-Seq assay for mammalian cells.
- Screened libraries of approximately 22,700 RNA sequences.
- Identified ribozyme devices responding to specific small molecules.
Main Results:
- Discovered new ribozyme devices responsive to theophylline, hypoxanthine, cyclic-di-GMP, and folinic acid.
- These devices exhibit low basal expression and high activation ratios.
- Identified conserved sequence and structure motifs for rational design.
Conclusions:
- The developed RNA-Seq assay enables efficient engineering of RNA devices in mammalian systems.
- Novel ribozyme switches significantly expand the toolkit for genetic device engineering.
- This approach advances the development of synthetic RNA genetic devices.
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