Engineered Streptomyces quorum-sensing components enable inducible siRNA-mediated translation control in mammalian cells and adjustable transcription control in mice

✍ Scribed by Wilfried Weber; Laetitia Malphettes; Maria de Jesus; Ronald Schoenmakers; Marie Daoud El-Baba; Manuela Spielmann; Bettina Keller; Cornelia C. Weber; Petra van de Wetering; Dominique Aubel; Florian M. Wurm; Martin Fussenegger

• Publisher: John Wiley and Sons
• Year: 2005
• Tongue: English
• Weight: 165 KB
• Volume: 7
• Category: Article
• ISSN: 1099-498X
• DOI: 10.1002/jgm.682

✦ Synopsis

Abstract

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. Copyright © 2004 John Wiley & Sons, Ltd.