Brief Communication abstract


Nature Methods 6, 63 - 66 (2008)
Published online: 30 November 2008 | doi:10.1038/nmeth.1277

Stable knockdown of microRNA in vivo by lentiviral vectors

Bernhard Gentner1, Giulia Schira1, Alice Giustacchini1,2, Mario Amendola1, Brian D Brown1,4, Maurilio Ponzoni2,3 & Luigi Naldini1,2

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Studying microRNA function in vivo requires genetic strategies to generate loss-of-function phenotypes. We used lentiviral vectors to stably and specifically knock down microRNA by overexpressing microRNA target sequences from polymerase II promoters. These vectors effectively inhibited regulation of reporter constructs and natural microRNA targets. We used bone marrow reconstitution with hematopoietic stem cells stably overexpressing miR-223 target sequence to phenocopy the genetic miR-223 knockout mouse, indicating robust interference of microRNA function in vivo.

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  1. San Raffaele Telethon Institute for Gene Therapy, Via Olgettina 58, 20132 Milan, Italy.
  2. Vita Salute San Raffaele University, Via Olgettina 58, 20132 Milan, Italy.
  3. Pathology Unit, San Raffaele Scientific Institute, Via Olgettina 58, 20132 Milan, Italy.
  4. Present address: Department of Genetics and Genomic Sciences, Mount Sinai School of Medicine, 1425 Madison Ave., New York, New York 10029, USA.

Correspondence to: Luigi Naldini1,2 e-mail: naldini.luigi@hsr.it



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