A synthetic transcription platform for programmable gene expression in mammalian cells.

Autor: Chen WCW; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. William.Chen@usd.edu.; Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. William.Chen@usd.edu.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. William.Chen@usd.edu.; Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA, 02114, USA. William.Chen@usd.edu.; Division of Basic Biomedical Sciences, Sanford School of Medicine, University of South Dakota, Vermillion, SD, 57069, USA. William.Chen@usd.edu., Gaidukov L; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Lai Y; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Wu MR; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA, 02215, USA., Cao J; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Gutbrod MJ; Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA, 02139, USA., Choi GCG; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Laboratory of Combinatorial Genetics and Synthetic Biology, School of Biomedical Sciences, The University of Hong Kong, Hong Kong, China., Utomo RP; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Department of Biochemistry, Wellesley College, Wellesley, MA, 02481, USA., Chen YC; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Taipei, Taiwan., Wroblewska L; Pfizer Inc., Cambridge, MA, 02139, USA., Kellis M; Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA, 02139, USA., Zhang L; Pfizer Inc., Andover, MA, 01810, USA., Weiss R; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Lu TK; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. tim@lugroup.org.; Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. tim@lugroup.org.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. tim@lugroup.org.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2022 Oct 18; Vol. 13 (1), pp. 6167. Date of Electronic Publication: 2022 Oct 18.
DOI: 10.1038/s41467-022-33287-9
Abstrakt: 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.
(© 2022. The Author(s).)
Databáze: MEDLINE
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