A synthetic transcription platform for programmable gene expression in mammalian cells.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
18 10 2022
18 10 2022
Historique:
received:
01
04
2021
accepted:
13
09
2022
entrez:
18
10
2022
pubmed:
19
10
2022
medline:
21
10
2022
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 36257931
doi: 10.1038/s41467-022-33287-9
pii: 10.1038/s41467-022-33287-9
pmc: PMC9579178
doi:
Substances chimiques
Interferon-gamma
82115-62-6
Transcription Factors
0
Antibodies, Monoclonal
0
Types de publication
Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6167Subventions
Organisme : NCI NIH HHS
ID : U01 CA250554
Pays : United States
Organisme : NHLBI NIH HHS
ID : T32 HL007208
Pays : United States
Informations de copyright
© 2022. The Author(s).
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