SEMPER: Stoichiometric expression of mRNA polycistrons by eukaryotic ribosomes for compact, ratio-tunable multi-gene expression
Abstract
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Acknowledgement
The authors thank Michael Elowitz for helpful discussions, James Linton for equipment training, and Eman Elsheikh and Arul Goel for experimental support. This work was supported by the National Institutes of Health (R01EB018975 to M.G.S.), the Millard and Muriel Jacobs Genetics and Genomics Laboratory at the California Institute of Technology, and the Flow Cytometry and Cell Sorting Facility at the California Institute of Technology. Related research in the Shapiro laboratory is supported by the Chan Zuckerberg Initiative. M.G.S. is an investigator of the Howard Hughes Medical Institute. BioRender.com
Contributions
M.D., I.D., and M.G.S. conceived and executed the study, analyzed data, and wrote the manuscript. M.D. and M.G.S. developed the initial hypothesis. M.D. and I.D. constructed the fluorescent protein SEMPER breadboarding system. M.D. performed and analyzed gas vesicle expression and ultrasound experiments as well as antibody expression and secretion experiments. A.L. prepared the IVT mRNA protocol. I.D. developed the SEMPER IVT mRNA breadboarding system. I.D. performed and analyzed IVT mRNA experiments and 2-ORF and 3-ORF FP expression experiments and developed the computational model. G.A. assisted in plasmid design and construction. M.Y.Y. assisted in the development of methionine-less FPs. M.D. and A.L. performed apoptosis assays.
Data Availability
Document S1. Figures S1–S13, Tables S1–S8, and supplemental references.
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Additional details
- R01EB018975
- National Institutes of Health
- Millard and Muriel Jacobs Genetics and Genomics Laboratory
- California Institute of Technology
- Chan Zuckerberg Initiative (United States)
- Howard Hughes Medical Institute
- Caltech groups
- Millard and Muriel Jacobs Genetics and Genomics Laboratory