Engineered Streptomyces quorum-sensing components enable inducible siRNA-mediated translation control in mammalian cells and adjustable transcription control in mice.
BACKGROUND
Recent advances in functional genomics, gene therapy, tissue engineering, drug discovery and biopharmaceuticals production have been fostered by precise small-molecule-mediated fine-tuning of desired transgenes.


METHODS
Capitalizing on well-evolved quorum-sensing regulatory networks in Streptomyces coelicolor we have designed a mammalian regulation system inducible by the non-toxic butyrolactone SCB1. Fusion of the S. coelicolor SCB1 quorum-sensing receptor ScbR to the human Kox-1-derived transsilencing domain reconstituted a mammalian transsilencer (SCS) able to repress transcription from SCS-specific operator-containing promoters in a reverse SCB1-adjustable manner.


RESULTS
This quorum-sensing-derived mammalian transgene control system (Q-ON) enabled precise SCB1-specific fine-tuning of (i) desired transgene transcription in a variety of mammalian/human cell lines and human primary cells, (ii) small interfering RNA-mediated posttranscriptional knockdown (siRNA) in mammalian cells, and (iii) dosing of a human glycoprotein in mice.


CONCLUSIONS
As exemplified by Q-ON technology, bacterial quorum-sensing regulons may represent a near-infinite source for the design of mammalian gene control systems compatible with molecular interventions relevant to future gene therapy and tissue engineering scenarios.
Weber
Wilfried
W
Institute of Biotechnology, Swiss Federal Institute of Technology, ETH Zurich, ETH Hoenggerberg, CH-8093 Zurich, Switzerland.
Malphettes
Laetitia
L
de Jesus
Maria
M
Schoenmakers
Ronald
R
El-Baba
Marie Daoud
MD
Spielmann
Manuela
M
Keller
Bettina
B
Weber
Cornelia C
CC
van de Wetering
Petra
P
Aubel
Dominique
D
Wurm
Florian M
FM
Fussenegger
Martin
M
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005786:Gene Expression Regulation / Q000187:drug effects; D005818:Genetic Engineering; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D014176:Protein Biosynthesis / Q000502:physiology*; D034741:RNA, Small Interfering / Q000502:physiology*; D013302:Streptomyces / Q000235:genetics*; D014158:Transcription, Genetic / Q000502:physiology*; D019076:Transgenes
D009841:Oligonucleotides; D034741:RNA, Small Interfering; C082868:butyrolactone I; D015107:4-Butyrolactone