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EVs containing Cre mRNA are sufficient to induce recombination in Purkinje neurons after intracerebellar injection in vivo.

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posted on 03.06.2014, 02:43 by Kirsten Ridder, Sascha Keller, Maria Dams, Anne-Kathleen Rupp, Jessica Schlaudraff, Domenico Del Turco, Julia Starmann, Jadranka Macas, Darja Karpova, Kavi Devraj, Candan Depboylu, Britta Landfried, Bernd Arnold, Karl H. Plate, Günter Höglinger, Holger Sültmann, Peter Altevogt, Stefan Momma

(A) EV preparations enriched for exosomes prepared from the peripheral blood and bone marrow of Vav-iCre mice were brought into the circulation by tail vein injection or were directly injected into the cerebellum. Injection of Cre RNA-containing EVs into tail veins did not lead to recombination events in the brain (n = 4). (B) β-galactosidase–positive Purkinje neuron in the cerebellum of a reporter mouse 4 d after intracerebellar injection of EVs. (C) Other reporter-gene–positive cells with a shape and size reminiscent of glial cells in proximity to the Purkinje cell layer. (D) Reporter-gene–positive cells displaying a microglia-like morphology. (E) Quantification of reporter-gene–expressing Purkinje neurons after intracerebellar injection of vesicle preparations from Vav-iCre–positive peripheral blood. Control mice (shaded part) were injected with 1 µl purified Cre-recombinase protein at 1 U/µl (light grey) or lysate prepared from Vav-iCre bone marrow (dark grey) and never showed any recombined cells. Scale bar, 50 µm (B and C) and 25 µm (D).