The platform has an expertise in the development, production and characterization of AAV and lentiviral vectors for the development of disease models and the evaluation of new treatment options (gene therapies and engineered cell therapies).

These viral vectors allow the stable or transient gene transfer in vitro in cell lines or primary cultures and in vivo in rodents or non-human primates. They can be used to over-express a gene of interest, inhibit the expression of an endogeneous gene (shRNA, CRISPR-Cas9), or express a reporter gene.​

Activities

• Development and production of virus vectors (lentiviruses and AAV). The quality control of the vectors includes titration by qPCR or ELISA, protein purity and sterility tests. After in vivo delivery, histology, TEP imaging and behavior platforms allow the assessment of the biodistribution and the biological effect of these vectors.​

• Biomarker assays in brain samples or cell culture extracts, neurodegenerative disease models (qPCR, ddPCR, electrophoresis and western blot, ELISA, enzymatic activity measurement, etc…).​

• Primary cell cultures (of neurons, astrocytes or mixed cultures) and cell lines. Time-lapse imaging analyses, toxicity tests (MTT, LDH, Calcein), flow cytometry.​

​ Laboratoire dédié aux études de biochimie et histologie (© Stroppa/CEA)

Laboratoire de culture cellulaire de biosécurité de niveau 3 (® Stroppa/CEA)

Equipement

• L1 laboratory for molecular biology and cellular microbiology dedicated to the cloning of genes of interest in eukaryotic expression plasmids, as well as to the study of gene expression in animal models. In addition to usual microbiology equipment, the laboratory counts with several qPCR thermal cyclers (96 and 384), a digital droplet PCR, a bioanalyzer and a nanodrop.​
  

• Well-equipped biosafety level 2 and 3 culture rooms (incubators, biosafety cabinets, binocular microscopes, inverted fluorescence microscopes including a time-lapse system). The L3 laboratory is composed of 10 rooms dedicated to the production and the handling of virus vectors.​

• L1 biochemistry laboratory dedicated to proteins analysis using various methods (Western blot, ELISA, etc…). It is equipped with a chemiluminescence detections system (Fusion FX7), fluorescence, microplate readers for fluorescence, luminescence and absorbance, and a cytometer (3 lasers).​