A synthetic transcription platform for programmable gene expression in mammalian cells
Abstract
Precise, scalable, and sustainable control of genetic and cellular activities in mammalian cells is key to developing precision therapeutics and smart biomanufacturing. Here we create a highly tunable, modular, versatile CRISPR-based synthetic transcription system for the programmable control of gene expression and cellular phenotypes in mammalian cells. Genetic circuits consisting of well-characterized libraries of guide RNAs, binding motifs of synthetic operators, transcriptional activators, and additional genetic regulatory elements express mammalian genes in a highly predictable and tunable manner. We demonstrate the programmable control of reporter genes episomally and chromosomally, with up to 25-fold more activity than seen with the EF1α promoter, in multiple cell types. We use these circuits to program the secretion of human monoclonal antibodies and to control T-cell effector function marked by interferon-γ production. Antibody titers and interferon-γ concentrations significantly correlate with synthetic promoter strengths, providing a platform for programming gene expression and cellular function in diverse applications.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Oct 18, 2022
- Source ID
- 10.1038/s41467-022-33287-9
Entities
People
- Chien-Wen Chen
- Gigi C.G. Choi
- Jicong Cao
- Leonid Gaidukov
- Liliana Wroblewska
- Lin Zhang
- Manolis Kellis
- Michael J. Gutbrod
- Ming-ru Wu
- Rachel P. Utomo
- Ron Weiss
- Timothy K. Lu
- Ying-chou Chen
- Yong Lai
Organizations
- National Heart, Lung, and Blood Institute
- United States Department of Defense
- University of South Dakota Sanford School of Medicine