Green fluorescent protein as a reporter for retrovirus and... : NeuroReport (original) (raw)

Neurochemistry

Green fluorescent protein as a reporter for retrovirus and helper virus-free HSV-1 amplicon vector-mediated gene transfer into neural cells in culture and in vivo

Aboody-Guterman, K S.1; Pechan, P A.1; Rainov, N G.1,5; Sena-Esteves, M1; Jacobs, A1; Snyder, E Y.2; Wild, P3; Schraner, E3; Tobler, K4,6; Breakefield, X O.1,7; Fraefel, C1

1Molecular Neurogenetics Unit, Neurology Service, Massachusetts General Hospital and Program in Neuroscience, Boston, MA, USA

2Department of Neurology and Pediatrics, The Children's Hospital, Harvard Medical School, Boston, MA, USA

3Department of Neurology and Pediatrics, Institute of Anatomy, Virology, University of Zurich, Zurich, Switzerland

4Department of Neurology and Pediatrics, Virology, University of Zurich, Zurich, Switzerland

5Department of Neurosurgery, Martin Luther University, Magdeburgerstrasse 16, D-06097 Halle, Germany;

6BMBCB/Northwestern University, 2153 North Campus Drive, Evanston, IL 60208, USA

7Corresponding Author: X.O. Breakefield

ACKNOWLEDGEMENTS: We thank Dr Francesca Persichetti for the preparation of primary cultures of rat striatal cells, Mr Sahar Nissim for the help with FACS analyses, and Dr Brian Seed, Dr David Jacoby, and Dr Howard Federoff for the kind gift of plasmid constructs.

This work was supported by the Harvard Medical School Lezler Award (KSA–G); the German Academy of Natural Sciences ‘Leopoldina’ (NGR); the Max- Planck–Society (AJ); NS33852 and NS34247, the Mental Retardation Research Center Grant HD18655, the Paralyzed Veterans of America, and the American Paralysis Association (EYS); NINDS NS24279, NCI CA69246, NIDCD DC002281, and the Brain Tumor Society (XOB); the Swiss National Science Foundation and the American Liver Foundation (CF).

Received 6 August 1997; accepted 18 September 1997

Abstract

GREEN fluorescent protein (GFP) is an effective marker for retrovirus and herpes virus vector-mediated gene transfer into various central nervous system-derived cells, both proliferative and non-proliferative, in culture and in vivo. Retrovirus vectors were used to stably transduce several rat and human glioma lines, and a multi-potent mouse neural progenitor line in culture. Implantation of selected pools of transduced glioma cells into rodent brain allowed clear visualization of the tumor and the invading tumor edge. Helper virus-free HSV-1 amplicon vectors successfully transferred gfp into non-dividing primary neural cells in culture and in the rat brain. This study describes the versatility of GFP for: (i) labelling of glioma cells in experimental brain tumor models and neural progenitor cells by retrovirus vectors, and (ii) efficient, non-toxic delivery of genes to post mitotic cells of the nervous system using helper-virus free HSV-1 amplicon vectors.