Article abstract
Nature Biotechnology 25, 1457 - 1467 (2007)
Published online: 16 November 2007 | doi:10.1038/nbt1372
Endogenous microRNA can be broadly exploited to regulate transgene expression according to tissue, lineage and differentiation state
Brian D Brown1,5,6, Bernhard Gentner1,6, Alessio Cantore1,2, Silvia Colleoni3,4, Mario Amendola1,2, Anna Zingale1, Alessia Baccarini1, Giovanna Lazzari3,4, Cesare Galli3,4 & Luigi Naldini1,2
Abstract
We have shown previously that transgene expression can be suppressed in hematopoietic cells using vectors that are responsive to microRNA (miRNA) regulation. Here we investigate the potential of this approach for more sophisticated control of transgene expression. Analysis of the relationship between miRNA expression levels and target mRNA suppression suggested that suppression depends on a threshold miRNA concentration. Using this information, we generated vectors that rapidly adjust transgene expression in response to changes in miRNA expression. These vectors sharply segregated transgene expression between closely related states of therapeutically relevant cells, including dendritic cells, hematopoietic and embryonic stem cells, and their progeny, allowing positive/negative selection according to the cells' differentiation state. Moreover, two miRNA target sites were combined to restrict transgene expression to a specific cell type in the liver. Notably, the vectors did not detectably perturb endogenous miRNA expression or regulation of natural targets. The properties of miRNA-regulated vectors should allow for safer and more effective therapeutic applications.
- San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, via Olgettina, 58, 20132 Milan, Italy.
- Vita Salute San Raffaele University, via Olgettina 58, 20132 Milan, Italy.
- Reproductive Technologies Laboratory, CIZ, Istituto Sperimentale Italiano Lazzaro Spallanzani, via Porcellasco 7/F, 26100 Cremona, Italy.
- Dipartimento Clinico Veterinario, University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy.
- Present address: Department of Genetics and Genomic Sciences, Mount Sinai School of Medicine, 1425 Madison Avenue, New York, New York 10029, USA.
- These authors contributed equally to the work.
Correspondence to: Luigi Naldini1,2 e-mail: naldini.luigi@hsr.it
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